Interaction Details


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
Fragment: 729-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: Fluorescein-KLPSTTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 25
Standard deviation of dissociation constant (10-6M): 6
Experiment method: FP_direct
Number of measurements: 3
Measured pKd: 4.6
PUBMED ID: 30726710

Experiment series 2

Protein A

Protein: RPS6KA1
Fragment: 726-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: Biotin-ado-ado-RVRKLPSTTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.14
Normalized holdup BI: 0.14
Immobilized partner concentration (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pKd: 3.96
PUBMED ID: 36115835
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

Experiment series 3

Protein A

Protein: RPS6KA1
Fragment: 683-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: ...KLPSTTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 185
Standard deviation of dissociation constant (10-6M): 25
Experiment method: FP_competitive
Number of measurements: 3
Measured pKd: 3.73
PUBMED ID: 30726710

Experiment series 4

Protein A

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPSTTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.11
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 31
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 4
Measured pKd: 3.61
PUBMED ID: 36115835

Experiment series 5

Protein A

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPSTTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.05
Normalized holdup BI: 0.05
Immobilized partner concentration (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pKd: 3.5
PUBMED ID: 36115835

Experiment series 6

Protein A

Protein: RPS6KA1
Fragment: 1-735
Construct: endogenous protein (Jurkat)

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Average holdup BI: 0.27
Immobilized partner concentration (10-6M): 10
P value: 2.58
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-08-03
Number of measurements: 3
The affinity was below the detection threshold of the assay.
PUBMED ID: 36542723
Experimental Details

-log10(P Two-sided unpaired T-test (6 vs 3))

Experiment series 7

Protein A

Protein: RPS6KA1
Fragment: 1-735
Construct: endogenous protein (Jurkat)

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Average holdup BI: 0.47
Immobilized partner concentration (10-6M): 10
P value: 1.37
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-06-27
Number of measurements: 6
The affinity was below the detection threshold of the assay.
PUBMED ID: 36542723
Experimental Details

-log10(P Two-sided unpaired T-test (6 vs 6))

Experiment series 8

Protein A

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: Biotin-ttds-RRVRKLPSTTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 0
Experiment method: SPR
Number of measurements: 1
The affinity was below the detection threshold of the assay.
PUBMED ID: 30726710

Experiment series 9

Protein A

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPSTTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 0
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835

Experiment series 10

Protein A

Protein: RPS6KA1
Fragment: 729-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: Fluorescein-KLP(pS)TTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
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
Number of measurements: 3
Measured pKd: 5.36
PUBMED ID: 30726710

Experiment series 11

Protein A

Protein: RPS6KA1
Fragment: 683-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: ...KLP(pS)TTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 32
Standard deviation of dissociation constant (10-6M): 5
Experiment method: FP_competitive
Number of measurements: 3
Measured pKd: 4.49
PUBMED ID: 30726710

Experiment series 12

Protein A

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.22
Normalized holdup BI: 0.25
Immobilized partner concentration (10-6M): 15
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pKd: 4.39
Experimental Details

not reported internal standards of holdup layout #5

Experiment series 13

Protein A

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: Biotin-ttds-RRVRKLP(pS)TTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 46
Standard deviation of dissociation constant (10-6M): 9
Experiment method: SPR
Number of measurements: 1
Measured pKd: 4.34
PUBMED ID: 30726710

Experiment series 14

Protein A

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.22
Normalized holdup BI: 0.23
Immobilized partner concentration (10-6M): 15
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pKd: 4.32
Experimental Details

not reported internal standards of holdup layout #5

Experiment series 15

Protein A

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.43
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 40
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 3
Measured pKd: 4.3
PUBMED ID: 36115835

Experiment series 16

Protein A

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
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
Number of measurements: 3
Measured pKd: 4.25
PUBMED ID: 36115835

Experiment series 17

Protein A

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.15
Normalized holdup BI: 0.15
Immobilized partner concentration (10-6M): 15
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pKd: 4.1
PUBMED ID: 36115835

Experiment series 18

Protein A

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (TPO)733
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPSpTTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.01
Normalized holdup BI: 0.01
Immobilized partner concentration (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835

Experiment series 19

Protein A

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (TPO)733
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPSpTTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 0
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835

Experiment series 20

Protein A

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (TPO)734
Sequence: RVRKLPSTTL
Construct: Biotin-ttds-RRVRKLPSTpTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
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 (10-6M): 25
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 2
Measured pKd: 3.92
PUBMED ID: 36115835

Experiment series 21

Protein A

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (TPO)734
Sequence: RVRKLPSTTL
Construct: Biotin-ttds-RRVRKLPSTpTL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 0
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835