Interaction Details


Details of experiments between Q96PY6 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: NEK1
Fragment: 1047-1056
Site: 14-3-3_pSer1052
Modification: (SEP)1052
Sequence: MLRTCSLPDL
Construct: MLRTC(pSer)LPDL (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: 0.01
Holdup BI standard deviation: 0.5
Immobilized partner concentration (10-6M): 130
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 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: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: -0.01
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 130
P value: 0.33
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: NEK1
Fragment: 280-289
Site: 14-3-3_pSer285
Modification: (SEP)285
Sequence: FSKFGSQPIP
Construct: FSKFG(pSer)QPIP (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: 0.09
Immobilized partner concentration (10-6M): 130
P value: 2.51
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.28
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: 294-303
Site: 14-3-3_pSer299
Modification: (SEP)299
Sequence: ASGQNSISVM
Construct: ASGQN(pSer)ISVM (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: 0.04
Immobilized partner concentration (10-6M): 130
P value: 0.78
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 5

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: 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: 0.06
Immobilized partner concentration (10-6M): 130
P value: 4.24
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 6

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: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: -0.02
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 130
P value: 0.54
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 7

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: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.02
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 130
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 8

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: 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: 0.06
Immobilized partner concentration (10-6M): 130
P value: 0.6
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))