Interaction Details


Details of experiments between Q8TEW0 and P62258
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.03
Holdup BI standard deviation: 0.47
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
P value: 0.05
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.76
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
P value: 5.77
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.34
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.64
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 116
P value: 5.55
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.19
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.03
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 116
P value: 1.19
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 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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.02
Holdup BI standard deviation: 0.07
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
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 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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.16
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
P value: 3.25
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.13
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.15
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 116
P value: 3.45
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 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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.2
Holdup BI standard deviation: 0.66
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
P value: 0.23
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.05
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 116
P value: 0.84
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.06
Holdup BI standard deviation: 0.08
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
P value: 0.81
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.25
Holdup BI standard deviation: 0.27
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 116
P value: 0.9
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.02
Holdup BI standard deviation: 0.33
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
P value: 0.05
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.41
Holdup BI standard deviation: 0.48
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 116
P value: 0.54
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.33
Holdup BI standard deviation: 0.25
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 116
P value: 1.72
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: 715-724
Site: 14-3-3_pSer720
Modification: (SEP)720
Sequence: SHSLYSGIEG
Construct: SHSLY(pSer)GIEG (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.04
Holdup BI standard deviation: 0.13
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 116
P value: 0.27
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 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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.03
Holdup BI standard deviation: 0.1
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
P value: 0.29
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 17

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.21
Holdup BI standard deviation: 0.23
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 116
P value: 0.93
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 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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.1
Holdup BI standard deviation: 0.06
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
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
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 19

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.04
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 116
P value: 0.89
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 20

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0
Holdup BI standard deviation: 0.07
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
P value: 0.01
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 21

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.29
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
P value: 5.95
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.47
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 22

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.24
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 116
P value: 5.17
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 23

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.26
Holdup BI standard deviation: 0.2
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 116
P value: 1.56
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 24

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.01
Holdup BI standard deviation: 0.09
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
P value: 0.11
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 25

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.36
Holdup BI standard deviation: 0.15
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
P value: 2.07
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.59
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 26

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.17
Holdup BI standard deviation: 0.23
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 116
P value: 0.7
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 27

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.46
Holdup BI standard deviation: 0.07
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
P value: 4.59
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.77
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 28

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.31
Holdup BI standard deviation: 0.06
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 116
P value: 4.04
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 29

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 116
P value: 0.07
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 30

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.05
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
P value: 2.08
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))