Interaction Details


Details of experiments between Q6R327 and P27348
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: RICTOR
Fragment: 1133-1142
Site: 14-3-3_pSer1138
Modification: (SEP)1138
Sequence: TLTEPSVDFN
Construct: TLTEP(pSer)VDFN (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: -0.1
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 130
P value: 1.42
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: RICTOR
Fragment: 1172-1181
Site: 14-3-3_pSer1177
Modification: (SEP)1177
Sequence: TGSTPSIGEN
Construct: TGSTP(pSer)IGEN (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: -0.2
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 130
P value: 2.53
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: RICTOR
Fragment: 1214-1223
Site: 14-3-3_pSer1219
Modification: (SEP)1219
Sequence: KIRSQSFNTD
Construct: biotin-ttds-KIRSQ(pSer)FNTD (HPLC-purified synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Measured dissociation constant (10-6M): 109
Standard deviation of dissociation constant (10-6M): 15
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-02-08
Number of measurements: 3
Measured pKd: 3.96
DOI: 10.1101/2022.12.08.519103

Experiment series 4

Protein A

Protein: RICTOR
Fragment: 1214-1223
Site: 14-3-3_pSer1219
Modification: (SEP)1219
Sequence: KIRSQSFNTD
Construct: KIRSQ(pSer)FNTD (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.53
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 130
P value: 5.47
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.93
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 5

Protein A

Protein: RICTOR
Fragment: 1279-1288
Site: 14-3-3_pSer1284
Modification: (SEP)1284
Sequence: LSKSNSVSLV
Construct: LSKSN(pSer)VSLV (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.37
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 130
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: 3.65
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: RICTOR
Fragment: 1297-1306
Site: 14-3-3_pSer1302
Modification: (SEP)1302
Sequence: PRRAQSLKAP
Construct: PRRAQ(pSer)LKAP (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.7
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 130
P value: 5.04
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.26
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: RICTOR
Fragment: 1380-1389
Site: 14-3-3_pSer1385
Modification: (SEP)1385
Sequence: FMKALSYASL
Construct: FMKAL(pSer)YASL (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.41
Holdup BI standard deviation: 0.21
Immobilized partner concentration (10-6M): 130
P value: 1.54
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.72
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: RICTOR
Fragment: 1383-1392
Site: 14-3-3_pSer1388
Modification: (SEP)1388
Sequence: ALSYASLDKE
Construct: ALSYA(pSer)LDKE (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 130
P value: 0.43
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: RICTOR
Fragment: 378-387
Site: 14-3-3_pSer383
Modification: (SEP)383
Sequence: PHRARSRPDL
Construct: PHRAR(pSer)RPDL (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.04
Holdup BI standard deviation: 0.21
Immobilized partner concentration (10-6M): 130
P value: 0.14
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 10

Protein A

Protein: RICTOR
Fragment: 1128-1137
Site: 14-3-3_pThr1133
Modification: (TPO)1133
Sequence: NRRIRTLTEP
Construct: NRRIR(pThr)LTEP (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.28
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 130
P value: 3.41
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.48
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: RICTOR
Fragment: 1130-1139
Site: 14-3-3_pThr1135
Modification: (TPO)1135
Sequence: RIRTLTEPSV
Construct: RIRTL(pThr)EPSV (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.27
Holdup BI standard deviation: 0.19
Immobilized partner concentration (10-6M): 130
P value: 1.49
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.46
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 12

Protein A

Protein: RICTOR
Fragment: 1327-1336
Site: 14-3-3_pThr1332
Modification: (TPO)1332
Sequence: AFGYATLKRL
Construct: AFGYA(pThr)LKRL (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: -0.25
Holdup BI standard deviation: 0.43
Immobilized partner concentration (10-6M): 130
P value: 0.68
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))