Interaction Details


Details of experiments between P31947 and Q6R327
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1079-1088
Site: 14-3-3_pSer1084
Modification: (SEP)1084
Sequence: IKDKNSFPFF
Construct: IKDKN(pSer)FPFF (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.38
Immobilized partner concentration (10-6M): 113
P value: 0.04
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 2

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1079-1088
Site: 14-3-3_pSer1084
Modification: (SEP)1084
Sequence: IKDKNSFPFF
Construct: IKDKN(pSer)FPFF (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.34
Immobilized partner concentration (10-6M): 142
P value: 0.26
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 3

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1079-1088
Site: 14-3-3_pSer1084
Modification: (SEP)1084
Sequence: IKDKNSFPFF
Construct: IKDKN(pSer)FPFF (crude synthetic peptide)
Average holdup BI: 0.31
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 111
P value: 2.21
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.61
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1133-1142
Site: 14-3-3_pSer1138
Modification: (SEP)1138
Sequence: TLTEPSVDFN
Construct: TLTEP(pSer)VDFN (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.25
Immobilized partner concentration (10-6M): 113
P value: 0.34
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1133-1142
Site: 14-3-3_pSer1138
Modification: (SEP)1138
Sequence: TLTEPSVDFN
Construct: TLTEP(pSer)VDFN (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 142
P value: 0.03
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1133-1142
Site: 14-3-3_pSer1138
Modification: (SEP)1138
Sequence: TLTEPSVDFN
Construct: TLTEP(pSer)VDFN (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 111
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 7

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1172-1181
Site: 14-3-3_pSer1177
Modification: (SEP)1177
Sequence: TGSTPSIGEN
Construct: TGSTP(pSer)IGEN (crude synthetic peptide)
Average holdup BI: -0.07
Holdup BI standard deviation: 0.1
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 8

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1172-1181
Site: 14-3-3_pSer1177
Modification: (SEP)1177
Sequence: TGSTPSIGEN
Construct: TGSTP(pSer)IGEN (crude synthetic peptide)
Average holdup BI: -0.12
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 142
P value: 2.22
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1172-1181
Site: 14-3-3_pSer1177
Modification: (SEP)1177
Sequence: TGSTPSIGEN
Construct: TGSTP(pSer)IGEN (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.16
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 10

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1214-1223
Site: 14-3-3_pSer1219
Modification: (SEP)1219
Sequence: KIRSQSFNTD
Construct: KIRSQ(pSer)FNTD (crude synthetic peptide)
Average holdup BI: 0.45
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 142
P value: 7.19
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 11

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1214-1223
Site: 14-3-3_pSer1219
Modification: (SEP)1219
Sequence: KIRSQSFNTD
Construct: KIRSQ(pSer)FNTD (crude synthetic peptide)
Average holdup BI: 0.36
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 6.03
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.7
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1214-1223
Site: 14-3-3_pSer1219
Modification: (SEP)1219
Sequence: KIRSQSFNTD
Construct: biotin-ttds-KIRSQ(pSer)FNTD (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 209
Standard deviation of dissociation constant (10-6M): 18
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-02-09
Number of measurements: 3
Measured pKd: 3.68
DOI: 10.1101/2022.12.08.519103

Experiment series 13

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1214-1223
Site: 14-3-3_pSer1219
Modification: (SEP)1219
Sequence: KIRSQSFNTD
Construct: KIRSQ(pSer)FNTD (crude synthetic peptide)
Average holdup BI: 0.3
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 111
P value: 3.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.58
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1279-1288
Site: 14-3-3_pSer1284
Modification: (SEP)1284
Sequence: LSKSNSVSLV
Construct: LSKSN(pSer)VSLV (crude synthetic peptide)
Average holdup BI: 0.22
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 5.8
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.39
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1279-1288
Site: 14-3-3_pSer1284
Modification: (SEP)1284
Sequence: LSKSNSVSLV
Construct: LSKSN(pSer)VSLV (crude synthetic peptide)
Average holdup BI: 0.25
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 4.95
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.38
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1279-1288
Site: 14-3-3_pSer1284
Modification: (SEP)1284
Sequence: LSKSNSVSLV
Construct: LSKSN(pSer)VSLV (crude synthetic peptide)
Average holdup BI: 0.27
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 4.89
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.51
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1297-1306
Site: 14-3-3_pSer1302
Modification: (SEP)1302
Sequence: PRRAQSLKAP
Construct: PRRAQ(pSer)LKAP (crude synthetic peptide)
Average holdup BI: 0.68
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 4.9
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.18
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1297-1306
Site: 14-3-3_pSer1302
Modification: (SEP)1302
Sequence: PRRAQSLKAP
Construct: PRRAQ(pSer)LKAP (crude synthetic peptide)
Average holdup BI: 0.5
Holdup BI standard deviation: 0.18
Immobilized partner concentration (10-6M): 113
P value: 2.66
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.95
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1297-1306
Site: 14-3-3_pSer1302
Modification: (SEP)1302
Sequence: PRRAQSLKAP
Construct: PRRAQ(pSer)LKAP (crude synthetic peptide)
Average holdup BI: 0.64
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 111
P value: 2.83
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.21
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1380-1389
Site: 14-3-3_pSer1385
Modification: (SEP)1385
Sequence: FMKALSYASL
Construct: FMKAL(pSer)YASL (crude synthetic peptide)
Average holdup BI: 0.39
Holdup BI standard deviation: 0.19
Immobilized partner concentration (10-6M): 113
P value: 2.11
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 21

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1380-1389
Site: 14-3-3_pSer1385
Modification: (SEP)1385
Sequence: FMKALSYASL
Construct: FMKAL(pSer)YASL (crude synthetic peptide)
Average holdup BI: 0.19
Holdup BI standard deviation: 0.31
Immobilized partner concentration (10-6M): 142
P value: 0.55
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 22

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1380-1389
Site: 14-3-3_pSer1385
Modification: (SEP)1385
Sequence: FMKALSYASL
Construct: FMKAL(pSer)YASL (crude synthetic peptide)
Average holdup BI: 0.35
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 111
P value: 2.23
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.69
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1383-1392
Site: 14-3-3_pSer1388
Modification: (SEP)1388
Sequence: ALSYASLDKE
Construct: ALSYA(pSer)LDKE (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 1.23
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1383-1392
Site: 14-3-3_pSer1388
Modification: (SEP)1388
Sequence: ALSYASLDKE
Construct: ALSYA(pSer)LDKE (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 142
P value: 0.04
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1383-1392
Site: 14-3-3_pSer1388
Modification: (SEP)1388
Sequence: ALSYASLDKE
Construct: ALSYA(pSer)LDKE (crude synthetic peptide)
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 26

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 378-387
Site: 14-3-3_pSer383
Modification: (SEP)383
Sequence: PHRARSRPDL
Construct: PHRAR(pSer)RPDL (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.21
Immobilized partner concentration (10-6M): 113
P value: 0.1
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 378-387
Site: 14-3-3_pSer383
Modification: (SEP)383
Sequence: PHRARSRPDL
Construct: PHRAR(pSer)RPDL (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.2
Immobilized partner concentration (10-6M): 142
P value: 0.18
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 28

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 378-387
Site: 14-3-3_pSer383
Modification: (SEP)383
Sequence: PHRARSRPDL
Construct: PHRAR(pSer)RPDL (crude synthetic peptide)
Average holdup BI: -0.23
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 111
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
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 29

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1128-1137
Site: 14-3-3_pThr1133
Modification: (TPO)1133
Sequence: NRRIRTLTEP
Construct: NRRIR(pThr)LTEP (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 113
P value: 0.76
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1128-1137
Site: 14-3-3_pThr1133
Modification: (TPO)1133
Sequence: NRRIRTLTEP
Construct: NRRIR(pThr)LTEP (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 142
P value: 1.83
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 31

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1128-1137
Site: 14-3-3_pThr1133
Modification: (TPO)1133
Sequence: NRRIRTLTEP
Construct: NRRIR(pThr)LTEP (crude synthetic peptide)
Average holdup BI: 0.25
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 111
P value: 2.53
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.48
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1130-1139
Site: 14-3-3_pThr1135
Modification: (TPO)1135
Sequence: RIRTLTEPSV
Construct: RIRTL(pThr)EPSV (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 113
P value: 0.41
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 33

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1130-1139
Site: 14-3-3_pThr1135
Modification: (TPO)1135
Sequence: RIRTLTEPSV
Construct: RIRTL(pThr)EPSV (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 142
P value: 0.91
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1130-1139
Site: 14-3-3_pThr1135
Modification: (TPO)1135
Sequence: RIRTLTEPSV
Construct: RIRTL(pThr)EPSV (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 111
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 35

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1327-1336
Site: 14-3-3_pThr1332
Modification: (TPO)1332
Sequence: AFGYATLKRL
Construct: AFGYA(pThr)LKRL (crude synthetic peptide)
Average holdup BI: -0.29
Holdup BI standard deviation: 0.37
Immobilized partner concentration (10-6M): 142
P value: 1.14
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 36

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RICTOR
Fragment: 1327-1336
Site: 14-3-3_pThr1332
Modification: (TPO)1332
Sequence: AFGYATLKRL
Construct: AFGYA(pThr)LKRL (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 111
P value: 0.41
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))