Details of experiments between Q6WCQ1 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: MPRIP
Protein A fragment: 296-305
Protein A site: 14-3-3_pSer301
Protein A modification: (SEP)301
Protein A sequence: PNHRRSQVIE
Protein A construct: PNHRR(pSer)QVIE (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Measured dissociation constant (10-6M): <100
Immobilized partner concentration in holdup experiment (10-6M): 115
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 1
Measured pKd: 4


Experiment series 2

Protein A protein: MPRIP
Protein A fragment: 988-997
Protein A site: 14-3-3_pSer993
Protein A modification: (SEP)993
Protein A sequence: IMKSKSNPDF
Protein A construct: IMKSK(pSer)NPDF (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Measured dissociation constant (10-6M): <10
Immobilized partner concentration in holdup experiment (10-6M): 115
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 1
Measured pKd: 5


Experiment series 3

Protein A protein: MPRIP
Protein A fragment: 988-997
Protein A site: 14-3-3_pSer993
Protein A modification: (SEP)993
Protein A sequence: IMKSKSNPDF
Protein A construct: IMKSK(pSer)NPDF (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Measured dissociation constant (10-6M): <1
Immobilized partner concentration in holdup experiment (10-6M): 129
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 1
Measured pKd: 6


Experiment series 4

Protein A protein: MPRIP
Protein A fragment: 1011-1020
Protein A site: 14-3-3_pSer1016
Protein A modification: (SEP)1016
Protein A sequence: NIRSKSVIEQ
Protein A construct: NIRSK(pSer)VIEQ (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.65
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 9.07
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 4.16


Experiment series 5

Protein A protein: MPRIP
Protein A fragment: 373-382
Protein A site: 14-3-3_pThr378
Protein A modification: (TPO)378
Protein A sequence: LDRRSTEPSV
Protein A construct: LDRRS(pThr)EPSV (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.75
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.71
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 4.37


Experiment series 6

Protein A protein: MPRIP
Protein A fragment: 537-546
Protein A site: 14-3-3_pThr542
Protein A modification: (TPO)542
Protein A sequence: EGRSKTFDWA
Protein A construct: EGRSK(pThr)FDWA (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.59
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.17
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 4.12


Experiment series 7

Protein A protein: MPRIP
Protein A fragment: 373-382
Protein A site: 14-3-3_pThr378
Protein A modification: (TPO)378
Protein A sequence: LDRRSTEPSV
Protein A construct: LDRRS(pThr)EPSV (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.73
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.76
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 4.31


Experiment series 8

Protein A protein: MPRIP
Protein A fragment: 537-546
Protein A site: 14-3-3_pThr542
Protein A modification: (TPO)542
Protein A sequence: EGRSKTFDWA
Protein A construct: EGRSK(pThr)FDWA (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.67
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.54
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 4.2


Experiment series 9

Protein A protein: MPRIP
Protein A fragment: 373-382
Protein A site: 14-3-3_pThr378
Protein A modification: (TPO)378
Protein A sequence: LDRRSTEPSV
Protein A construct: LDRRS(pThr)EPSV (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.63
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.29
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 4.18


Experiment series 10

Protein A protein: MPRIP
Protein A fragment: 537-546
Protein A site: 14-3-3_pThr542
Protein A modification: (TPO)542
Protein A sequence: EGRSKTFDWA
Protein A construct: EGRSK(pThr)FDWA (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.75
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.03
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 4.35


Experiment series 11

Protein A protein: MPRIP
Protein A fragment: 289-298
Protein A site: 14-3-3_pSer294
Protein A modification: (SEP)294
Protein A sequence: SPPSPSTPNH
Protein A construct: SPPSP(pSer)TPNH (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.24
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.98
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 12

Protein A protein: MPRIP
Protein A fragment: 1011-1020
Protein A site: 14-3-3_pSer1016
Protein A modification: (SEP)1016
Protein A sequence: NIRSKSVIEQ
Protein A construct: NIRSK(pSer)VIEQ (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.58
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.41
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 4.09


Experiment series 13

Protein A protein: MPRIP
Protein A fragment: 1011-1020
Protein A site: 14-3-3_pSer1016
Protein A modification: (SEP)1016
Protein A sequence: NIRSKSVIEQ
Protein A construct: NIRSK(pSer)VIEQ (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.56
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.99
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 3.97


Experiment series 14

Protein A protein: MPRIP
Protein A fragment: 264-273
Protein A site: 14-3-3_pSer269
Protein A modification: (SEP)269
Protein A sequence: ESGYFSLEKT
Protein A construct: ESGYF(pSer)LEKT (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.16
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.57
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 15

Protein A protein: MPRIP
Protein A fragment: 988-997
Protein A site: 14-3-3_pSer993
Protein A modification: (SEP)993
Protein A sequence: IMKSKSNPDF
Protein A construct: IMKSK(pSer)NPDF (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 1
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.92
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 6.43


Experiment series 16

Protein A protein: MPRIP
Protein A fragment: 264-273
Protein A site: 14-3-3_pSer269
Protein A modification: (SEP)269
Protein A sequence: ESGYFSLEKT
Protein A construct: ESGYF(pSer)LEKT (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.22
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.84
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 17

Protein A protein: MPRIP
Protein A fragment: 289-298
Protein A site: 14-3-3_pSer294
Protein A modification: (SEP)294
Protein A sequence: SPPSPSTPNH
Protein A construct: SPPSP(pSer)TPNH (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.22
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.7
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 18

Protein A protein: MPRIP
Protein A fragment: 264-273
Protein A site: 14-3-3_pSer269
Protein A modification: (SEP)269
Protein A sequence: ESGYFSLEKT
Protein A construct: ESGYF(pSer)LEKT (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.17
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.32
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 19

Protein A protein: MPRIP
Protein A fragment: 289-298
Protein A site: 14-3-3_pSer294
Protein A modification: (SEP)294
Protein A sequence: SPPSPSTPNH
Protein A construct: SPPSP(pSer)TPNH (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.16
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.04
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 20

Protein A protein: MPRIP
Protein A fragment: 296-305
Protein A site: 14-3-3_pSer301
Protein A modification: (SEP)301
Protein A sequence: PNHRRSQVIE
Protein A construct: PNHRR(pSer)QVIE (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: -0.33
Holdup BI standard deviation: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 21

Protein A protein: MPRIP
Protein A fragment: 296-305
Protein A site: 14-3-3_pSer301
Protein A modification: (SEP)301
Protein A sequence: PNHRRSQVIE
Protein A construct: PNHRR(pSer)QVIE (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.2
Holdup BI standard deviation: 0.44
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.26
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 22

Protein A protein: MPRIP
Protein A fragment: 1011-1020
Protein A site: 14-3-3_pSer1016
Protein A modification: (SEP)1016
Protein A sequence: NIRSKSLKEG
Protein A construct: NIRSK(pSer)LKEG (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: -0.22
Holdup BI standard deviation: 1.16
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.15
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.


Experiment series 23

Protein A protein: MPRIP
Protein A fragment: 1011-1020
Protein A site: 14-3-3_pSer1016
Protein A modification: (SEP)1016
Protein A sequence: NIRSKSLKEG
Protein A construct: NIRSK(pSer)LKEG (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.03
Holdup BI standard deviation: 0.71
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.02
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.