Interaction Details


Details of experiments between Q8TEW0 and P31947
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: PARD3
Fragment: 1173-1182
Site: 14-3-3_pSer1178
Modification: (SEP)1178
Sequence: RRRTYSFEQP
Construct: RRRTY(pSer)FEQP (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.21
Holdup BI standard deviation: 0.53
Immobilized partner concentration (10-6M): 142
P value: 0.41
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 2

Protein A

Protein: PARD3
Fragment: 1218-1227
Site: 14-3-3_pSer1223
Modification: (SEP)1223
Sequence: QRQYSSLPRQ
Construct: QRQYS(pSer)LPRQ (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.5
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 4.55
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.85
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 3

Protein A

Protein: PARD3
Fragment: 1218-1227
Site: 14-3-3_pSer1223
Modification: (SEP)1223
Sequence: QRQYSSLPRQ
Construct: QRQYS(pSer)LPRQ (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.38
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 113
P value: 4.6
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.74
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 4

Protein A

Protein: PARD3
Fragment: 1332-1341
Site: 14-3-3_pSer1337
Modification: (SEP)1337
Sequence: VPPSPSQVAR
Construct: VPPSP(pSer)QVAR (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.04
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 2.04
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 5

Protein A

Protein: PARD3
Fragment: 1332-1341
Site: 14-3-3_pSer1337
Modification: (SEP)1337
Sequence: VPPSPSQVAR
Construct: VPPSP(pSer)QVAR (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.02
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 142
P value: 0.3
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 6

Protein A

Protein: PARD3
Fragment: 1351-1360
Site: 14-3-3_pSer1356
Modification: (SEP)1356
Sequence: GRPFYS----
Construct: TPEKGRPFY(pSer) (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.15
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 113
P value: 2.85
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 7

Protein A

Protein: PARD3
Fragment: 1351-1360
Site: 14-3-3_pSer1356
Modification: (SEP)1356
Sequence: GRPFYS----
Construct: TPEKGRPFY(pSer) (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.13
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 3.15
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 8

Protein A

Protein: PARD3
Fragment: 139-148
Site: 14-3-3_pSer144
Modification: (SEP)144
Sequence: HVRRSSDPAL
Construct: HVRRS(pSer)DPAL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.19
Holdup BI standard deviation: 0.46
Immobilized partner concentration (10-6M): 142
P value: 0.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 9

Protein A

Protein: PARD3
Fragment: 196-205
Site: 14-3-3_pSer201
Modification: (SEP)201
Sequence: DENYRSLPRD
Construct: DENYR(pSer)LPRD (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.02
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 113
P value: 0.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 10

Protein A

Protein: PARD3
Fragment: 196-205
Site: 14-3-3_pSer201
Modification: (SEP)201
Sequence: DENYRSLPRD
Construct: DENYR(pSer)LPRD (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.04
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 0.51
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 11

Protein A

Protein: PARD3
Fragment: 216-225
Site: 14-3-3_pSer221
Modification: (SEP)221
Sequence: DNARSSLSAS
Construct: DNARS(pSer)LSAS (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.06
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 113
P value: 0.49
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 12

Protein A

Protein: PARD3
Fragment: 216-225
Site: 14-3-3_pSer221
Modification: (SEP)221
Sequence: DNARSSLSAS
Construct: DNARS(pSer)LSAS (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.05
Holdup BI standard deviation: 0.2
Immobilized partner concentration (10-6M): 142
P value: 0.25
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 13

Protein A

Protein: PARD3
Fragment: 20-29
Site: 14-3-3_pSer25
Modification: (SEP)25
Sequence: HMKVFSLIQQ
Construct: HMKVF(pSer)LIQQ (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.16
Holdup BI standard deviation: 0.27
Immobilized partner concentration (10-6M): 113
P value: 0.37
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 14

Protein A

Protein: PARD3
Fragment: 710-719
Site: 14-3-3_pSer715
Modification: (SEP)715
Sequence: RERRISHSLY
Construct: RERRI(pSer)HSLY (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.49
Holdup BI standard deviation: 0.35
Immobilized partner concentration (10-6M): 113
P value: 1.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 15

Protein A

Protein: PARD3
Fragment: 710-719
Site: 14-3-3_pSer715
Modification: (SEP)715
Sequence: RERRISHSLY
Construct: RERRI(pSer)HSLY (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.21
Holdup BI standard deviation: 0.26
Immobilized partner concentration (10-6M): 142
P value: 0.79
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 16

Protein A

Protein: PARD3
Fragment: 715-724
Site: 14-3-3_pSer720
Modification: (SEP)720
Sequence: SHSLYSGIEG
Construct: SHSLY(pSer)GIEG (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.08
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 113
P value: 0.52
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 17

Protein A

Protein: PARD3
Fragment: 715-724
Site: 14-3-3_pSer720
Modification: (SEP)720
Sequence: SHSLYSGIEG
Construct: SHSLY(pSer)GIEG (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.03
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 0.38
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 18

Protein A

Protein: PARD3
Fragment: 822-831
Site: 14-3-3_pSer827
Modification: (SEP)827
Sequence: GFGRQSMSEK
Construct: GFGRQ(pSer)MSEK (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.15
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 113
P value: 0.83
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 19

Protein A

Protein: PARD3
Fragment: 822-831
Site: 14-3-3_pSer827
Modification: (SEP)827
Sequence: GFGRQSMSEK
Construct: GFGRQ(pSer)MSEK (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.07
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
P value: 1.17
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 20

Protein A

Protein: PARD3
Fragment: 847-856
Site: 14-3-3_pSer852
Modification: (SEP)852
Sequence: TRKSKSMDLG
Construct: TRKSK(pSer)MDLG (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Measured dissociation constant (10-6M): <100
Immobilized partner concentration (10-6M): 113
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 1
Measured pKd: 4
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 21

Protein A

Protein: PARD3
Fragment: 876-885
Site: 14-3-3_pSer881
Modification: (SEP)881
Sequence: RDVGPSLGLK
Construct: RDVGP(pSer)LGLK (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.01
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 0.14
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 22

Protein A

Protein: PARD3
Fragment: 876-885
Site: 14-3-3_pSer881
Modification: (SEP)881
Sequence: RDVGPSLGLK
Construct: RDVGP(pSer)LGLK (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.01
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 0.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 23

Protein A

Protein: PARD3
Fragment: 884-893
Site: 14-3-3_pSer889
Modification: (SEP)889
Sequence: LKKSSSLESL
Construct: LKKSS(pSer)LESL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.4
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 142
P value: 6.6
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.67
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 24

Protein A

Protein: PARD3
Fragment: 884-893
Site: 14-3-3_pSer889
Modification: (SEP)889
Sequence: LKKSSSLESL
Construct: LKKSS(pSer)LESL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.31
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 5.77
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.59
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 25

Protein A

Protein: PARD3
Fragment: 968-977
Site: 14-3-3_pSer973
Modification: (SEP)973
Sequence: DQPSHSLERQ
Construct: DQPSH(pSer)LERQ (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.14
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 113
P value: 1.13
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 26

Protein A

Protein: PARD3
Fragment: 968-977
Site: 14-3-3_pSer973
Modification: (SEP)973
Sequence: DQPSHSLERQ
Construct: DQPSH(pSer)LERQ (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.01
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 142
P value: 0.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 27

Protein A

Protein: PARD3
Fragment: 171-180
Site: 14-3-3_pThr176
Modification: (TPO)176
Sequence: TRWSTTAGFL
Construct: TRWST(pThr)AGFL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.31
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 113
P value: 1.66
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.59
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 28

Protein A

Protein: PARD3
Fragment: 171-180
Site: 14-3-3_pThr176
Modification: (TPO)176
Sequence: TRWSTTAGFL
Construct: TRWST(pThr)AGFL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.34
Holdup BI standard deviation: 0.18
Immobilized partner concentration (10-6M): 142
P value: 1.75
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.55
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 29

Protein A

Protein: PARD3
Fragment: 293-302
Site: 14-3-3_pThr298
Modification: (TPO)298
Sequence: ARGGRTLGLL
Construct: ARGGR(pThr)LGLL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.34
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 142
P value: 3.67
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.56
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 30

Protein A

Protein: PARD3
Fragment: 293-302
Site: 14-3-3_pThr298
Modification: (TPO)298
Sequence: ARGGRTLGLL
Construct: ARGGR(pThr)LGLL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.17
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 113
P value: 2.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 31

Protein A

Protein: PARD3
Fragment: 476-485
Site: 14-3-3_pThr481
Modification: (TPO)481
Sequence: TSRDVTIGGS
Construct: TSRDV(pThr)IGGS (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.03
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 113
P value: 2.92
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 32

Protein A

Protein: PARD3
Fragment: 476-485
Site: 14-3-3_pThr481
Modification: (TPO)481
Sequence: TSRDVTIGGS
Construct: TSRDV(pThr)IGGS (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.04
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 142
P value: 1.52
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 33

Protein A

Protein: PARD3
Fragment: 828-837
Site: 14-3-3_pThr833
Modification: (TPO)833
Sequence: MSEKRTKQFS
Construct: MSEKR(pThr)KQFS (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.03
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 142
P value: 0.27
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 34

Protein A

Protein: PARD3
Fragment: 1218-1227
Site: 14-3-3_pSer1223
Modification: (SEP)1223
Sequence: QRQYSSLPRQ
Construct: QRQYS(pSer)LPRQ (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.4
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 111
P value: 3.94
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.78
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 35

Protein A

Protein: PARD3
Fragment: 1332-1341
Site: 14-3-3_pSer1337
Modification: (SEP)1337
Sequence: VPPSPSQVAR
Construct: VPPSP(pSer)QVAR (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
P value: 0.17
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 36

Protein A

Protein: PARD3
Fragment: 1351-1360
Site: 14-3-3_pSer1356
Modification: (SEP)1356
Sequence: GRPFYS----
Construct: TPEKGRPFY(pSer) (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.13
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 111
P value: 2.04
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 37

Protein A

Protein: PARD3
Fragment: 139-148
Site: 14-3-3_pSer144
Modification: (SEP)144
Sequence: HVRRSSDPAL
Construct: HVRRS(pSer)DPAL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0
Holdup BI standard deviation: 0.27
Immobilized partner concentration (10-6M): 111
P value: 0
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 38

Protein A

Protein: PARD3
Fragment: 196-205
Site: 14-3-3_pSer201
Modification: (SEP)201
Sequence: DENYRSLPRD
Construct: DENYR(pSer)LPRD (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
P value: 0.11
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 39

Protein A

Protein: PARD3
Fragment: 216-225
Site: 14-3-3_pSer221
Modification: (SEP)221
Sequence: DNARSSLSAS
Construct: DNARS(pSer)LSAS (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.46
Holdup BI standard deviation: 1.21
Immobilized partner concentration (10-6M): 111
P value: 0.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 40

Protein A

Protein: PARD3
Fragment: 20-29
Site: 14-3-3_pSer25
Modification: (SEP)25
Sequence: HMKVFSLIQQ
Construct: HMKVF(pSer)LIQQ (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.46
Holdup BI standard deviation: 0.77
Immobilized partner concentration (10-6M): 111
P value: 0.86
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 41

Protein A

Protein: PARD3
Fragment: 710-719
Site: 14-3-3_pSer715
Modification: (SEP)715
Sequence: RERRISHSLY
Construct: RERRI(pSer)HSLY (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.07
Holdup BI standard deviation: 0.43
Immobilized partner concentration (10-6M): 111
P value: 0.11
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 42

Protein A

Protein: PARD3
Fragment: 715-724
Site: 14-3-3_pSer720
Modification: (SEP)720
Sequence: SHSLYSGIEG
Construct: SHSLY(pSer)GIEG (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.02
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 111
P value: 0.13
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 43

Protein A

Protein: PARD3
Fragment: 822-831
Site: 14-3-3_pSer827
Modification: (SEP)827
Sequence: GFGRQSMSEK
Construct: GFGRQ(pSer)MSEK (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.1
Holdup BI standard deviation: 0.45
Immobilized partner concentration (10-6M): 111
P value: 0.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 44

Protein A

Protein: PARD3
Fragment: 876-885
Site: 14-3-3_pSer881
Modification: (SEP)881
Sequence: RDVGPSLGLK
Construct: RDVGP(pSer)LGLK (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
P value: 0.15
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 45

Protein A

Protein: PARD3
Fragment: 884-893
Site: 14-3-3_pSer889
Modification: (SEP)889
Sequence: LKKSSSLESL
Construct: LKKSS(pSer)LESL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.29
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 111
P value: 2.71
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.56
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 46

Protein A

Protein: PARD3
Fragment: 968-977
Site: 14-3-3_pSer973
Modification: (SEP)973
Sequence: DQPSHSLERQ
Construct: DQPSH(pSer)LERQ (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.01
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 111
P value: 0.05
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 47

Protein A

Protein: PARD3
Fragment: 171-180
Site: 14-3-3_pThr176
Modification: (TPO)176
Sequence: TRWSTTAGFL
Construct: TRWST(pThr)AGFL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.46
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 111
P value: 3.37
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.88
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 48

Protein A

Protein: PARD3
Fragment: 293-302
Site: 14-3-3_pThr298
Modification: (TPO)298
Sequence: ARGGRTLGLL
Construct: ARGGR(pThr)LGLL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.38
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 111
P value: 3.94
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.75
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 49

Protein A

Protein: PARD3
Fragment: 476-485
Site: 14-3-3_pThr481
Modification: (TPO)481
Sequence: TSRDVTIGGS
Construct: TSRDV(pThr)IGGS (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.02
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 0.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))