Details of experiments between Q5SQI0 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: ATAT1
Protein A fragment: 1-421
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: 0.74
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): 3.68
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: 5.46


Experiment series 2

Protein A protein: ATAT1
Protein A fragment: 210-224
Protein A site: PxxP_1
Protein A sequence: AAVDPTPAAPARKLP
Protein A construct: Bio Ado Ado-AAVDPTPAAPARKLP-CONH2

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

Average holdup BI: 0.08
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: 3.72


Experiment series 3

Protein A protein: ATAT1
Protein A fragment: 293-307
Protein A site: PxxP_2
Protein A sequence: RSLRLCPPHPTARLL
Protein A construct: Bio Ado Ado-RSLRLCPPHPTARLL-CONH2

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

Average holdup BI: 0.05
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: 3.51