Details of experiments between Q15418 and Q9H5P4
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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.13
Holdup BI standard deviation: 0.05
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: 3.56


Experiment series 2

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: PDZD7
Protein B fragment: 199-313
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.11
Holdup BI standard deviation: 0.2
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: 3.49


Experiment series 3

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: PDZD7
Protein B fragment: 82-170
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.09
Holdup BI standard deviation: 0.09
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: 3
The affinity was below the detection threshold of the assay.


Experiment series 4

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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.64
Holdup BI standard deviation: 0.06
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: 5
Measured pKd: 4.79


Experiment series 5

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: PDZD7
Protein B fragment: 82-170
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.02
Holdup BI standard deviation: 0.03
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: 4
The affinity was below the detection threshold of the assay.


Experiment series 6

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: PDZD7
Protein B fragment: 199-313
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0
Holdup BI standard deviation: 0.11
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: 5
The affinity was below the detection threshold of the assay.


Experiment series 7

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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 11.9
Standard deviation of dissociation constant (10-6M): 2.31
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.92


Experiment series 8

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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 99.41
Standard deviation of dissociation constant (10-6M): 16.92
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


Experiment series 9

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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 0
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
The affinity was below the detection threshold of the assay.


Experiment series 10

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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 12.7
Standard deviation of dissociation constant (10-6M): 2.52
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.9


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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.06
Normalized holdup BI: 0.07
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
Measured pKd: 3.72


Experiment series 12

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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.04
Normalized holdup BI: 0.05
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: 3.5


Experiment series 13

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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.54
Normalized holdup BI: 0.65
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: 1
Measured pKd: 4.91


Experiment series 14

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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.46
Normalized holdup BI: 0.57
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: 4.93


Experiment series 15

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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.17
Normalized holdup BI: 0.2
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: 4.24


Experiment series 16

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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.6
Normalized holdup BI: 0.63
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: 5.04


Experiment series 17

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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 0.8
Standard deviation of dissociation constant (10-6M): 0.05
Experiment method: FP_direct
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 6.1


Experiment series 18

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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 1.8
Standard deviation of dissociation constant (10-6M): 0.1
Experiment method: FP_direct
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 5.74


Experiment series 19

Protein A protein: RPS6KA1
Protein A fragment: 683-735
Protein A site: PBM
Protein A sequence: RVRKLPSTTL
Protein A construct: ...KLPSTTL

Protein B protein: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 4
Standard deviation of dissociation constant (10-6M): 1
Experiment method: FP_competitive
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 5.4


Experiment series 20

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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 46
Standard deviation of dissociation constant (10-6M): 7
Experiment method: FP_competitive
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.34


Experiment series 21

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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 6.2
Standard deviation of dissociation constant (10-6M): 0.9
Experiment method: SPR
PUBMED ID of publication: 30726710
Number of measurements: 1
Measured pKd: 5.21


Experiment series 22

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: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
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.