Details of experiments between Q5JSZ5 and P49418
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A protein: PRRC2B
Protein A fragment: 1-2229
Protein A construct: endogenous protein (Jurkat)

Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3

Average holdup BI: 0.85
Immobilized partner concentration in holdup experiment (10-6M): 10
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.34
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.06.29.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
Number of measurements: 3
Measured pKd: 5.76

Experiment series 2

Protein A protein: PRRC2B
Protein A fragment: 251-265
Protein A site: PxxP_1
Protein A sequence: KDPSLRPAQPVRKGA
Protein A construct: Bio Ado Ado-KDPSLRPAQPVRKGA-CONH2

Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.

Experiment series 3

Protein A protein: PRRC2B
Protein A fragment: 306-320
Protein A site: PxxP_2
Protein A sequence: PQLRLEPRVPFRQFQ
Protein A construct: Bio Ado Ado-PQLRLEPRVPFRQFQ-CONH2

Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.18
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 2
Measured pKd: 4.11

Experiment series 4

Protein A protein: PRRC2B
Protein A fragment: 330-344
Protein A site: PxxP_3
Protein A sequence: RLGLSRPLRPLRQLV
Protein A construct: Bio Ado Ado-RLGLSRPLRPLRQLV-CONH2

Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.7
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 5.18

Experiment series 5

Protein A protein: PRRC2B
Protein A fragment: 445-459
Protein A site: PxxP_4
Protein A sequence: VRKAPDPQPPPRKLH
Protein A construct: Bio Ado Ado-VRKAPDPQPPPRKLH-CONH2

Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.45
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 4.71

Experiment series 6

Protein A protein: PRRC2B
Protein A fragment: 548-562
Protein A site: PxxP_5
Protein A sequence: QAEKEVPWSPSAEKA
Protein A construct: Bio Ado Ado-QAEKEVPWSPSAEKA-CONH2

Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.06
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.

Experiment series 7

Protein A protein: PRRC2B
Protein A fragment: 570-584
Protein A site: PxxP_6
Protein A sequence: AVHKGSPEFPAQETP
Protein A construct: Bio Ado Ado-AVHKGSPEFPAQETP-CONH2

Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.

Experiment series 8

Protein A protein: PRRC2B
Protein A fragment: 647-661
Protein A site: PxxP_7
Protein A sequence: QPVYPPPSHPQRTFY
Protein A construct: Bio Ado Ado-QPVYPPPSHPQRTFY-CONH2

Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 3.78

Experiment series 9

Protein A protein: PRRC2B
Protein A fragment: 848-862
Protein A site: PxxP_8
Protein A sequence: QNLRCSPLEPDFVPD
Protein A construct: Bio Ado Ado-QNLRCSPLEPDFVPD-CONH2

Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: -0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.