Details of experiments between Q15418 and Q96L92
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPpSTTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.43
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 40
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.3


Experiment series 2

Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPSTTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.11
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 31
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 4
Measured pKd: 3.61


Experiment series 3

Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPSTTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 0
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
The affinity was below the detection threshold of the assay.


Experiment series 4

Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPpSTTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 56.51
Standard deviation of dissociation constant (10-6M): 3.88
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.25


Experiment series 5

Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (TPO)733
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPSpTTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 0
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
The affinity was below the detection threshold of the assay.


Experiment series 6

Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (TPO)734
Protein A sequence: RVRKLPSTTL
Protein A construct: Biotin-ttds-RRVRKLPSTpTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 0
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
The affinity was below the detection threshold of the assay.


Experiment series 7

Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPpSTTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.15
Normalized holdup BI: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 15
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.1


Experiment series 8

Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (TPO)733
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPSpTTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.01
Normalized holdup BI: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 9

Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (TPO)734
Protein A sequence: RVRKLPSTTL
Protein A construct: Biotin-ttds-RRVRKLPSTpTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.15
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.17
Normalized holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 25
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
PUBMED ID of publication: 36115835
Number of measurements: 2
Measured pKd: 3.92


Experiment series 10

Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPSTTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.05
Normalized holdup BI: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.5


Experiment series 11

Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPpSTTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.22
Normalized holdup BI: 0.25
Immobilized partner concentration in holdup experiment (10-6M): 15
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: not reported internal standards of holdup layout #5
Number of measurements: 1
Measured pKd: 4.39


Experiment series 12

Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPpSTTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.22
Normalized holdup BI: 0.23
Immobilized partner concentration in holdup experiment (10-6M): 15
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: not reported internal standards of holdup layout #5
Number of measurements: 1
Measured pKd: 4.32


Experiment series 13

Protein A protein: RPS6KA1
Protein A fragment: 726-735
Protein A site: PBM
Protein A sequence: RVRKLPSTTL
Protein A construct: Biotin-ado-ado-RVRKLPSTTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.14
Normalized holdup BI: 0.14
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.96


Experiment series 14

Protein A protein: RPS6KA1
Protein A fragment: 729-735
Protein A site: PBM
Protein A sequence: RVRKLPSTTL
Protein A construct: Fluorescein-KLPSTTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 25
Standard deviation of dissociation constant (10-6M): 6
Experiment method: FP_direct
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.6


Experiment series 15

Protein A protein: RPS6KA1
Protein A fragment: 729-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: Fluorescein-KLP(pS)TTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 4.4
Standard deviation of dissociation constant (10-6M): 0.4
Experiment method: FP_direct
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 5.36


Experiment series 16

Protein A protein: RPS6KA1
Protein A fragment: 683-735
Protein A site: PBM
Protein A sequence: RVRKLPSTTL
Protein A construct: ...KLPSTTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 185
Standard deviation of dissociation constant (10-6M): 25
Experiment method: FP_competitive
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 3.73


Experiment series 17

Protein A protein: RPS6KA1
Protein A fragment: 683-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: ...KLP(pS)TTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 32
Standard deviation of dissociation constant (10-6M): 5
Experiment method: FP_competitive
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.49


Experiment series 18

Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A sequence: RVRKLPSTTL
Protein A construct: Biotin-ttds-RRVRKLPSTTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 0
Experiment method: SPR
PUBMED ID of publication: 30726710
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 19

Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: Biotin-ttds-RRVRKLP(pS)TTL

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 46
Standard deviation of dissociation constant (10-6M): 9
Experiment method: SPR
PUBMED ID of publication: 30726710
Number of measurements: 1
Measured pKd: 4.34


Experiment series 20

Protein A protein: RPS6KA1
Protein A fragment: 1-735
Protein A construct: endogenous protein (Jurkat)

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: avi(biotin)-his6-MBP-TEVsite-PDZ

Average holdup BI: 0.47
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): 1.37
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.06.27.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 6))
PUBMED ID of publication: 36542723
Number of measurements: 6
The affinity was below the detection threshold of the assay.


Experiment series 21

Protein A protein: RPS6KA1
Protein A fragment: 1-735
Protein A construct: endogenous protein (Jurkat)

Protein B protein: SNX27
Protein B fragment: 40-141
Protein B site: PDZ
Protein B construct: avi(biotin)-his6-MBP-TEVsite-PDZ

Average holdup BI: 0.27
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): 2.58
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.08.03.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
PUBMED ID of publication: 36542723
Number of measurements: 3
The affinity was below the detection threshold of the assay.