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
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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.33
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: 5
Measured pKd: 4.1


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: PARD3B
Protein B fragment: 376-471
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.03
Holdup BI standard deviation: 0.04
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 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: PARD3B
Protein B fragment: 490-594
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.08
Holdup BI standard deviation: 0.03
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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.43
Holdup BI standard deviation: 0.14
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: 4.41


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: PARD3B
Protein B fragment: 376-471
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: -0.01
Holdup BI standard deviation: 0.05
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: PARD3B
Protein B fragment: 490-594
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: -0.03
Holdup BI standard deviation: 0.02
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 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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B 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

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.31


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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B 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

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.72


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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B 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

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 3.99


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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B 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

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 3.9


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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B 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 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: 2
Measured pKd: 4.62


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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B 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 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: 2
Measured pKd: 3.9


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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.21
Normalized holdup BI: 0.21
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: 4.04


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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B 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 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: 2
Measured pKd: 4.4


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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.4
Normalized holdup BI: 0.4
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: 4.69


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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.45
Normalized holdup BI: 0.45
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: 4.7


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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 27
Standard deviation of dissociation constant (10-6M): 3
Experiment method: FP_direct
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.57


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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B 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
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 5.17


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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 45
Standard deviation of dissociation constant (10-6M): 7
Experiment method: FP_competitive
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.35


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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 31
Standard deviation of dissociation constant (10-6M): 3.5
Experiment method: FP_competitive
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.51


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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 4
Standard deviation of dissociation constant (10-6M): 0.3
Experiment method: SPR
PUBMED ID of publication: 30726710
Number of measurements: 1
Measured pKd: 5.4


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: PARD3B
Protein B fragment: 184-299
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 6.1
Standard deviation of dissociation constant (10-6M): 0.4
Experiment method: SPR
PUBMED ID of publication: 30726710
Number of measurements: 1
Measured pKd: 5.21