Details of experiments between Q92974 and Q04917
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: ARHGEF2
Protein A fragment: 881-890
Protein A site: 14-3-3_pSer886
Protein A modification: (SEP)886
Protein A sequence: DPRRRSLPAG
Protein A construct: DPRRR(pSer)LPAG (crude synthetic peptide)

Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta

Average holdup BI: 0.74
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.77
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 4.33


Experiment series 2

Protein A protein: ARHGEF2
Protein A fragment: 104-113
Protein A site: 14-3-3_pSer109
Protein A modification: (SEP)109
Protein A sequence: ALQSVSLRSK
Protein A construct: ALQSV(pSer)LRSK (crude synthetic peptide)

Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta

Average holdup BI: 0.59
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.35
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 4.04


Experiment series 3

Protein A protein: ARHGEF2
Protein A fragment: 117-126
Protein A site: 14-3-3_pSer122
Protein A modification: (SEP)122
Protein A sequence: RERPSSAIYP
Protein A construct: RERPS(pSer)AIYP (crude synthetic peptide)

Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta

Average holdup BI: 0.5
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.13
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 3.88


Experiment series 4

Protein A protein: ARHGEF2
Protein A fragment: 181-190
Protein A site: 14-3-3_pSer186
Protein A modification: (SEP)186
Protein A sequence: RNRTLSVESL
Protein A construct: RNRTL(pSer)VESL (crude synthetic peptide)

Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta

Average holdup BI: 0.42
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.36
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 3.75


Experiment series 5

Protein A protein: ARHGEF2
Protein A fragment: 146-155
Protein A site: 14-3-3_pSer151
Protein A modification: (SEP)151
Protein A sequence: LAKSVSTTNI
Protein A construct: LAKSV(pSer)TTNI (crude synthetic peptide)

Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta

Average holdup BI: 0.63
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.29
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 4.11


Experiment series 6

Protein A protein: ARHGEF2
Protein A fragment: 640-649
Protein A site: 14-3-3_pSer645
Protein A modification: (SEP)645
Protein A sequence: LFRSESLESP
Protein A construct: LFRSE(pSer)LESP (crude synthetic peptide)

Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta

Average holdup BI: 0.22
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.93
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 3.33


Experiment series 7

Protein A protein: ARHGEF2
Protein A fragment: 167-176
Protein A site: 14-3-3_pSer172
Protein A modification: (SEP)172
Protein A sequence: LRRILSQSTD
Protein A construct: LRRIL(pSer)QSTD (crude synthetic peptide)

Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta

Average holdup BI: 0.18
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.54
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 8

Protein A protein: ARHGEF2
Protein A fragment: 128-137
Protein A site: 14-3-3_pSer133
Protein A modification: (SEP)133
Protein A sequence: DSFRQSLLGS
Protein A construct: DSFRQ(pSer)LLGS (crude synthetic peptide)

Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta

Average holdup BI: 0.03
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.56
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 9

Protein A protein: ARHGEF2
Protein A fragment: 948-957
Protein A site: 14-3-3_pSer953
Protein A modification: (SEP)953
Protein A sequence: EEEGSSRLSP
Protein A construct: EEEGS(pSer)RLSP (crude synthetic peptide)

Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta

Average holdup BI: -0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 10

Protein A protein: ARHGEF2
Protein A fragment: 936-945
Protein A site: 14-3-3_pSer941
Protein A modification: (SEP)941
Protein A sequence: RLQDSSDPDT
Protein A construct: RLQDS(pSer)DPDT (crude synthetic peptide)

Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta

Average holdup BI: -0.16
Holdup BI standard deviation: 0.45
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.38
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 11

Protein A protein: ARHGEF2
Protein A fragment: 172-181
Protein A site: 14-3-3_pSer177
Protein A modification: (SEP)177
Protein A sequence: SQSTDSLNMR
Protein A construct: SQSTD(pSer)LNMR (crude synthetic peptide)

Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta

Average holdup BI: 0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.24
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 12

Protein A protein: ARHGEF2
Protein A fragment: 706-715
Protein A site: 14-3-3_pSer711
Protein A modification: (SEP)711
Protein A sequence: RTFNGSIEL-
Protein A construct: ARTFNG(pSer)IEL (crude synthetic peptide)

Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta

Average holdup BI: -0.07
Holdup BI standard deviation: 0.31
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.21
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 13

Protein A protein: ARHGEF2
Protein A fragment: 138-147
Protein A site: 14-3-3_pSer143
Protein A modification: (SEP)143
Protein A sequence: RRGRSSLSLA
Protein A construct: RRGRS(pSer)LSLA (crude synthetic peptide)

Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta

Average holdup BI: 0.01
Holdup BI standard deviation: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.07
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 14

Protein A protein: ARHGEF2
Protein A fragment: 124-133
Protein A site: 14-3-3_pSer129
Protein A modification: (SEP)129
Protein A sequence: IYPSDSFRQS
Protein A construct: IYPSD(pSer)FRQS (crude synthetic peptide)

Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta

Average holdup BI: 0
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.07
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.