Interaction Details


Details of experiments between Q15418 and Q9NZN5
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: 726-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: Biotin-ado-ado-RVRKLPSTTL

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.67
Normalized holdup BI: 0.89
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: 5.75
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 2

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 2.79
Standard deviation of dissociation constant (10-6M): 0.11
Experiment method: SPR
Number of measurements: 1
Measured pKd: 5.55
PUBMED ID: 30726710

Experiment series 3

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.73
Normalized holdup BI: 0.81
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: 5.45
PUBMED ID: 36115835

Experiment series 4

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 6.6
Standard deviation of dissociation constant (10-6M): 1.7
Experiment method: FP_competitive
Number of measurements: 3
Measured pKd: 5.18
PUBMED ID: 30726710

Experiment series 5

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 6.96
Standard deviation of dissociation constant (10-6M): 0.71
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 5.16
PUBMED ID: 36115835

Experiment series 6

Protein A

Protein: RPS6KA1
Fragment: 729-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: Fluorescein-KLPSTTL

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 7.5
Standard deviation of dissociation constant (10-6M): 0.8
Experiment method: FP_direct
Number of measurements: 3
Measured pKd: 5.12
PUBMED ID: 30726710

Experiment series 7

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.76
Holdup BI standard deviation: 0.03
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: 8
Measured pKd: 5.06
PUBMED ID: 36115835

Experiment series 8

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 15
Experiment method: ITC
Number of measurements: 1
Measured pKd: 4.82
PUBMED ID: 30726710

Experiment series 9

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 29
Standard deviation of dissociation constant (10-6M): 8
Experiment method: FP_direct
Number of measurements: 3
Measured pKd: 4.54
PUBMED ID: 30726710

Experiment series 10

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 52
Experiment method: ITC
Number of measurements: 1
Measured pKd: 4.28
PUBMED ID: 30726710

Experiment series 11

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.11
Normalized holdup BI: 0.14
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: 4.05
Experimental Details

not reported internal standards of holdup layout #5

Experiment series 12

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 98.42
Standard deviation of dissociation constant (10-6M): 11.17
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 4.01
PUBMED ID: 36115835

Experiment series 13

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.08
Normalized holdup BI: 0.08
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: 3.79
PUBMED ID: 36115835

Experiment series 14

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.14
Holdup BI standard deviation: 0.12
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: 5
Measured pKd: 3.61
PUBMED ID: 36115835

Experiment series 15

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 0
Experiment method: FP_competitive
Number of measurements: 3
The affinity was below the detection threshold of the assay.
PUBMED ID: 30726710

Experiment series 16

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
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 17

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.04
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
The affinity was below the detection threshold of the assay.
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: ARHGEF12
Fragment: 67-151
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 19

Protein A

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

Protein B

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 13.26
Standard deviation of dissociation constant (10-6M): 1.47
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 4.88
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: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.48
Normalized holdup BI: 0.62
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: 1
Measured pKd: 4.87
PUBMED ID: 36115835