Details of experiments between Q15418 and Q92743
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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.16
Holdup BI standard deviation: 0.18
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: 4
Measured pKd: 3.69


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.41
Holdup BI standard deviation: 0.09
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: 8
Measured pKd: 4.38


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 68.13
Standard deviation of dissociation constant (10-6M): 10.04
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.17


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 78.73
Standard deviation of dissociation constant (10-6M): 10.62
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.1


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 131.3
Standard deviation of dissociation constant (10-6M): 10.9
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 3.88


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 30.73
Standard deviation of dissociation constant (10-6M): 2.39
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.51


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.02
Normalized holdup BI: 0.03
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
The affinity was below the detection threshold of the assay.


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.05
Normalized holdup BI: 0.07
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: 3.65


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.41
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.53
Normalized holdup BI standard deviation: 0.02
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: 4.7


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.12
Normalized holdup BI: 0.17
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: 4.09


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.09
Normalized holdup BI: 0.12
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: 3.99


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.12
Normalized holdup BI: 0.14
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: 4.07


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.16
Normalized holdup BI: 0.2
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: 4.16


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 30
Standard deviation of dissociation constant (10-6M): 2
Experiment method: FP_direct
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.52


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 11.3
Standard deviation of dissociation constant (10-6M): 0.4
Experiment method: FP_direct
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.95


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 19
Standard deviation of dissociation constant (10-6M): 3
Experiment method: FP_competitive
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.72


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: HTRA1
Protein B fragment: 365-480
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 33
Standard deviation of dissociation constant (10-6M): 2
Experiment method: FP_competitive
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.48


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: HTRA1
Protein B fragment: 365-480
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: HTRA1
Protein B fragment: 365-480
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.