Details of experiments between P61978 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: HNRNPK
Protein A fragment: 1-463
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: 0.11
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): 4.48
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
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: HNRNPK
Protein A fragment: 266-280
Protein A site: PxxP_1
Protein A sequence: PGRGGRPMPPSRRDY
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.35
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.51
Experiment series 3
Protein A protein: HNRNPK
Protein A fragment: 283-297
Protein A site: PxxP_2
Protein A sequence: MSPRRGPPPPPPGRG
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.23
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.25
Experiment series 4
Protein A protein: HNRNPK
Protein A fragment: 284-298
Protein A site: PxxP_3
Protein A sequence: SPRRGPPPPPPGRGG
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.26
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.32
Experiment series 5
Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.28
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.37
Experiment series 6
Protein A protein: HNRNPK
Protein A fragment: 302-316
Protein A site: PxxP_5
Protein A sequence: SRARNLPLPPPPPPR
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.29
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.39
Experiment series 7
Protein A protein: HNRNPK
Protein A fragment: 305-319
Protein A site: PxxP_6
Protein A sequence: RNLPLPPPPPPRGGD
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.1
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 3.78
Experiment series 8
Protein A protein: HNRNPK
Protein A fragment: 285-299
Protein A site: PxxP_4
Protein A sequence: PRRGPPPPPPGRGGR
Protein A construct: Bio E E Ado Ado-PRRGPPPPPPGRGGR-CONH2
Protein B protein: AMPH
Protein B fragment: 615-695
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.34
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: 4.49