Interaction Details


Details of experiments between Q96PY6 and P61981
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: NEK1
Fragment: 1047-1056
Site: 14-3-3_pSer1052
Modification: (SEP)1052
Sequence: MLRTCSLPDL
Construct: MLRTC(pSer)LPDL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.2
Holdup BI standard deviation: 0.46
Immobilized partner concentration (10-6M): 129
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 2

Protein A

Protein: NEK1
Fragment: 9-18
Site: 14-3-3_pSer14
Modification: (SEP)14
Sequence: KIGEGSFGKA
Construct: KIGEG(pSer)FGKA (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.01
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 0.25
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 3

Protein A

Protein: NEK1
Fragment: 9-18
Site: 14-3-3_pSer14
Modification: (SEP)14
Sequence: KIGEGSFGKA
Construct: KIGEG(pSer)FGKA (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.01
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 129
P value: 0.37
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 4

Protein A

Protein: NEK1
Fragment: 280-289
Site: 14-3-3_pSer285
Modification: (SEP)285
Sequence: FSKFGSQPIP
Construct: FSKFG(pSer)QPIP (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.54
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 134
P value: 5.31
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.94
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: NEK1
Fragment: 280-289
Site: 14-3-3_pSer285
Modification: (SEP)285
Sequence: FSKFGSQPIP
Construct: FSKFG(pSer)QPIP (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.48
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 129
P value: 4.79
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.86
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: NEK1
Fragment: 294-303
Site: 14-3-3_pSer299
Modification: (SEP)299
Sequence: ASGQNSISVM
Construct: ASGQN(pSer)ISVM (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.16
Holdup BI standard deviation: 0.61
Immobilized partner concentration (10-6M): 134
P value: 0.2
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: NEK1
Fragment: 294-303
Site: 14-3-3_pSer299
Modification: (SEP)299
Sequence: ASGQNSISVM
Construct: ASGQN(pSer)ISVM (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.06
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 129
P value: 0.86
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: NEK1
Fragment: 409-418
Site: 14-3-3_pSer414
Modification: (SEP)414
Sequence: WRNVLSAGGS
Construct: WRNVL(pSer)AGGS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.51
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 5.12
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.89
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 9

Protein A

Protein: NEK1
Fragment: 409-418
Site: 14-3-3_pSer414
Modification: (SEP)414
Sequence: WRNVLSAGGS
Construct: WRNVL(pSer)AGGS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.48
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 129
P value: 5.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.86
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: NEK1
Fragment: 413-422
Site: 14-3-3_pSer418
Modification: (SEP)418
Sequence: LSAGGSGEVK
Construct: LSAGG(pSer)GEVK (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.08
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 134
P value: 1.3
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 11

Protein A

Protein: NEK1
Fragment: 413-422
Site: 14-3-3_pSer418
Modification: (SEP)418
Sequence: LSAGGSGEVK
Construct: LSAGG(pSer)GEVK (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.02
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 129
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 12

Protein A

Protein: NEK1
Fragment: 136-145
Site: 14-3-3_pThr141
Modification: (TPO)141
Sequence: LTKDGTVQLG
Construct: LTKDG(pThr)VQLG (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.08
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 134
P value: 1.64
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 13

Protein A

Protein: NEK1
Fragment: 136-145
Site: 14-3-3_pThr141
Modification: (TPO)141
Sequence: LTKDGTVQLG
Construct: LTKDG(pThr)VQLG (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 129
P value: 0.07
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 14

Protein A

Protein: NEK1
Fragment: 151-160
Site: 14-3-3_pThr156
Modification: (TPO)156
Sequence: RVLNSTVELA
Construct: RVLNS(pThr)VELA (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.05
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 134
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 15

Protein A

Protein: NEK1
Fragment: 151-160
Site: 14-3-3_pThr156
Modification: (TPO)156
Sequence: RVLNSTVELA
Construct: RVLNS(pThr)VELA (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.1
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 129
P value: 1.79
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: NEK1
Fragment: 1047-1056
Site: 14-3-3_pSer1052
Modification: (SEP)1052
Sequence: MLRTCSLPDL
Construct: MLRTC(pSer)LPDL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.16
Holdup BI standard deviation: 0.25
Immobilized partner concentration (10-6M): 115
P value: 0.61
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: NEK1
Fragment: 1121-1130
Site: 14-3-3_pSer1126
Modification: (SEP)1126
Sequence: PGEEYSEEEE
Construct: PGEEY(pSer)EEEE (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.26
Holdup BI standard deviation: 1.88
Immobilized partner concentration (10-6M): 115
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 18

Protein A

Protein: NEK1
Fragment: 9-18
Site: 14-3-3_pSer14
Modification: (SEP)14
Sequence: KIGEGSFGKA
Construct: KIGEG(pSer)FGKA (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.03
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 115
P value: 0.63
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: NEK1
Fragment: 280-289
Site: 14-3-3_pSer285
Modification: (SEP)285
Sequence: FSKFGSQPIP
Construct: FSKFG(pSer)QPIP (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.39
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 115
P value: 2.49
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.77
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: NEK1
Fragment: 294-303
Site: 14-3-3_pSer299
Modification: (SEP)299
Sequence: ASGQNSISVM
Construct: ASGQN(pSer)ISVM (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.28
Holdup BI standard deviation: 0.75
Immobilized partner concentration (10-6M): 115
P value: 0.32
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 21

Protein A

Protein: NEK1
Fragment: 409-418
Site: 14-3-3_pSer414
Modification: (SEP)414
Sequence: WRNVLSAGGS
Construct: WRNVL(pSer)AGGS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.42
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 115
P value: 5.75
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.81
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: NEK1
Fragment: 413-422
Site: 14-3-3_pSer418
Modification: (SEP)418
Sequence: LSAGGSGEVK
Construct: LSAGG(pSer)GEVK (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.03
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 115
P value: 0.22
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: NEK1
Fragment: 136-145
Site: 14-3-3_pThr141
Modification: (TPO)141
Sequence: LTKDGTVQLG
Construct: LTKDG(pThr)VQLG (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.05
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 115
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 24

Protein A

Protein: NEK1
Fragment: 151-160
Site: 14-3-3_pThr156
Modification: (TPO)156
Sequence: RVLNSTVELA
Construct: RVLNS(pThr)VELA (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.12
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 115
P value: 2.24
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))