Interaction Details


Details of experiments between Q15418 and Q9Y2H9
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: 683-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: ...KLPSTTL

Protein B

Protein: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 5
Standard deviation of dissociation constant (10-6M): 1
Experiment method: FP_competitive
Number of measurements: 3
Measured pKd: 5.3
PUBMED ID: 30726710

Experiment series 2

Protein A

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

Protein B

Protein: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 9.38
Standard deviation of dissociation constant (10-6M): 0.64
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 5.03
PUBMED ID: 36115835

Experiment series 3

Protein A

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

Protein B

Protein: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.6
Holdup BI standard deviation: 0.06
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: 4.72
PUBMED ID: 36115835

Experiment series 4

Protein A

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

Protein B

Protein: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 20
Standard deviation of dissociation constant (10-6M): 1
Experiment method: SPR
Number of measurements: 1
Measured pKd: 4.7
PUBMED ID: 30726710

Experiment series 5

Protein A

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

Protein B

Protein: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.22
Normalized holdup BI: 0.44
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: 4.69
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 6

Protein A

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

Protein B

Protein: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.22
Normalized holdup BI: 0.44
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: 4.68
PUBMED ID: 36115835

Experiment series 7

Protein A

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

Protein B

Protein: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 26
Standard deviation of dissociation constant (10-6M): 4
Experiment method: FP_direct
Number of measurements: 3
Measured pKd: 4.59
PUBMED ID: 30726710

Experiment series 8

Protein A

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

Protein B

Protein: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 34
Standard deviation of dissociation constant (10-6M): 8
Experiment method: FP_direct
Number of measurements: 3
Measured pKd: 4.47
PUBMED ID: 30726710

Experiment series 9

Protein A

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

Protein B

Protein: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 85.92
Standard deviation of dissociation constant (10-6M): 7.28
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 4.07
PUBMED ID: 36115835

Experiment series 10

Protein A

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

Protein B

Protein: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 92
Standard deviation of dissociation constant (10-6M): 12
Experiment method: FP_competitive
Number of measurements: 3
Measured pKd: 4.04
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: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.06
Normalized holdup BI: 0.13
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: 3.99
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: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0
Normalized holdup BI: 0.01
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
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835

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: MAST1
Fragment: 965-1056
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 14

Protein A

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

Protein B

Protein: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.07
Holdup BI standard deviation: 0.08
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
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835

Experiment series 15

Protein A

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

Protein B

Protein: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.01
Normalized holdup BI: 0.03
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 16

Protein A

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

Protein B

Protein: MAST1
Fragment: 965-1056
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 17

Protein A

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

Protein B

Protein: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 10.55
Standard deviation of dissociation constant (10-6M): 0.21
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 4.98
PUBMED ID: 36115835

Experiment series 18

Protein A

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

Protein B

Protein: MAST1
Fragment: 965-1056
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.19
Normalized holdup BI: 0.43
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: 4.5
PUBMED ID: 36115835