Interaction Details


Details of experiments between Q15418 and Q8TEW8
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: 725-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: Biotin-ttds-RRVRKLPSTTL

Protein B

Protein: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 4
Standard deviation of dissociation constant (10-6M): 0.3
Experiment method: SPR
Number of measurements: 1
Measured pKd: 5.4
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: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.45
Normalized holdup BI: 0.45
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: 4.7
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: 729-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: Fluorescein-KLPSTTL

Protein B

Protein: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 27
Standard deviation of dissociation constant (10-6M): 3
Experiment method: FP_direct
Number of measurements: 3
Measured pKd: 4.57
PUBMED ID: 30726710

Experiment series 4

Protein A

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

Protein B

Protein: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.43
Holdup BI standard deviation: 0.14
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: 5
Measured pKd: 4.41
PUBMED ID: 36115835

Experiment series 5

Protein A

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

Protein B

Protein: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.24
Holdup BI standard deviation: 0
Normalized holdup BI: 0.3
Normalized holdup BI standard deviation: 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: 2
Measured pKd: 4.4
PUBMED ID: 36115835

Experiment series 6

Protein A

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

Protein B

Protein: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 45
Standard deviation of dissociation constant (10-6M): 7
Experiment method: FP_competitive
Number of measurements: 3
Measured pKd: 4.35
PUBMED ID: 30726710

Experiment series 7

Protein A

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

Protein B

Protein: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 49.11
Standard deviation of dissociation constant (10-6M): 2.37
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 4.31
PUBMED ID: 36115835

Experiment series 8

Protein A

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

Protein B

Protein: PARD3B
Fragment: 376-471
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.01
Holdup BI standard deviation: 0.05
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
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835

Experiment series 9

Protein A

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

Protein B

Protein: PARD3B
Fragment: 490-594
Site: PDZ_3
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.03
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
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835

Experiment series 10

Protein A

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

Protein B

Protein: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 6.1
Standard deviation of dissociation constant (10-6M): 0.4
Experiment method: SPR
Number of measurements: 1
Measured pKd: 5.21
PUBMED ID: 30726710

Experiment series 11

Protein A

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

Protein B

Protein: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 6.8
Standard deviation of dissociation constant (10-6M): 0.5
Experiment method: FP_direct
Number of measurements: 3
Measured pKd: 5.17
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: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 19.16
Standard deviation of dissociation constant (10-6M): 0.95
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 4.72
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: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.4
Normalized holdup BI: 0.4
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: 4.69
Experimental Details

not reported internal standards of holdup layout #5

Experiment series 14

Protein A

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

Protein B

Protein: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.29
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.36
Normalized holdup BI standard deviation: 0.04
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: 2
Measured pKd: 4.62
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: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 31
Standard deviation of dissociation constant (10-6M): 3.5
Experiment method: FP_competitive
Number of measurements: 3
Measured pKd: 4.51
PUBMED ID: 30726710

Experiment series 16

Protein A

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

Protein B

Protein: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.33
Holdup BI standard deviation: 0.09
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: 4.1
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: PARD3B
Fragment: 376-471
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.03
Holdup BI standard deviation: 0.04
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
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: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Protein B

Protein: PARD3B
Fragment: 490-594
Site: PDZ_3
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.08
Holdup BI standard deviation: 0.03
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
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: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 101.55
Standard deviation of dissociation constant (10-6M): 6.69
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 3.99
PUBMED ID: 36115835

Experiment series 20

Protein A

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

Protein B

Protein: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.1
Holdup BI standard deviation: 0
Normalized holdup BI: 0.12
Normalized holdup BI standard deviation: 0
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: 2
Measured pKd: 3.9
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: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.21
Normalized holdup BI: 0.21
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: 4.04
PUBMED ID: 36115835

Experiment series 22

Protein A

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

Protein B

Protein: PARD3B
Fragment: 184-299
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 126.96
Standard deviation of dissociation constant (10-6M): 11.49
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 3.9
PUBMED ID: 36115835