This codebook.txt file was generated on 20200423 by wilsonkm

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GENERAL INFORMATION
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1. Title of Dataset 
 Analysis of the kinetics of binding of Protein Kinase A Inhibitor alpha (PKIa) to cAMP-dependent protein kinase a catalytic subunit (PKA-C) 

2. Author Information

  Principal Investigator Contact Information
        Name: Gianluigi Veglia
           Institution: University of Minnesota
           Address: 
           Email: vegli001@umn.edu


3. Date of data collection: 2015/12-2016/02

4. Geographic location of data collection : Biophysical Technology Center, University of Minnesota

5. Information about funding sources that supported the collection of the data:
Sponsorship: NIH GM 100310 to G.V

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SHARING/ACCESS INFORMATION
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1. Licenses/restrictions placed on the data:
CC0 1.0 Universal

2. Links to publications that cite or use the data: 
Olivieri, C., Wang, Y., Li, G. C., Subrahmanian, M. V., Kim, J., Stultz, B. R., ... & Gao, J. (2020). Multi-state recognition pathway of the intrinsically disordered protein kinase inhibitor by protein kinase A. Elife, 9, e55607.
10.7554/eLife.55607


3. Recommended citation for the data: 
 Li, Geoffrey; Muretta, Joseph; Olivieri, Cristina. (2020). Analysis of the kinetics of binding of Protein Kinase A Inhibitor alpha (PKIa) to cAMP-dependent protein kinase a catalytic subunit (PKA-C). Retrieved from the Data Repository for the University of Minnesota, http://hdl.handle.net/11299/212383. 
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DATA & FILE OVERVIEW
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1. File List
	A. Filename: SteadyState_Fluo_PKA+PKI_FRET.xlsx
	Short description:    Raw TR-fret data of the titrations of acceptor-label-PKI to Saturate ATP PKA-C complex(donor-labeled).
	Three different cys mutants of PKIa have been used (V3C, S28C, S59C). All these mutants were label with the FRET acceptor tetramethylrhodamine-5-maleimide (TMR) 
	A cys mutant of PKA-C has been used (C199A,S325C) and  labeled with the FRET Donor Alexa Fluor 488.
	
        
	B. Filename:  TR-FRET_PKI-PKA.xlsx           
	Short description: Raw TR-fret data of the titrations of PKA-C to double-labeled PKI  
 	PKA-C C199A,S325C mutant without fluorescent label was titrated to a double mutant of PKIa with two cysteines at residue 3 and 59 (PKIV3C,S59C). the
	PKIV3C,S59C mutant that was labeled with both Alexa-488 (donor) and TMR (acceptor) at positions 3
	and 59, respectively (PKIDONOR-ACCEPTOR). 

2. Relationship between files:        
	These data has been used to calculate kinetic parameter of the binding event of PKI to PKA-C and PKA-C to PKI. 

3. Additional related data collected that was not included in the current data package: NO


4. Are there multiple versions of the dataset? no

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METHODOLOGICAL INFORMATION
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1. Description of methods used for collection/generation of data: 
	Time-Resolved FRET is a combination of time-resolved fluorometry (TRF) with Förster resonance energy transfer (FRET) that allows the determination of kintecits parameters with incresed flexibility, reliability and sensitivity compared to the individual FRET and TRF experiments.
	Muretta JM et al, "Direct real-time detection of the structural and biochemical events in the myosin power stroke", PNAS November 17, 2015 112 (46) 14272-14277, https://doi.org/10.1073/pnas.1514859112
	Nesmelov YE et al, "Structural kinetics of myosin by transient time-resolved FRET", PNAS February 1, 2011 108 (5) 1891-1896; https://doi.org/10.1073/pnas.1012320108
	
2. Methods for processing the data: 
	The total fluorescence was fit into equations describing either single, double or triple exponential decays. More detailed information of the data analysis are described in the Material and Methods section of the eLife pubblication.

3. Instrument- or software-specific information needed to interpret the data:
	Both data set were acquired using an home-built stopped-flow spectrofluorometer at Biophysical Technology Center (University of Minnesota)
	Data analyses were done using Origin 8 (OriginLab) software.


4. Environmental/experimental conditions:
	All the experiment were recorded at 25˚C using the following buffer: 20 mM KH2PO4, 90 mM KCl, 10 mM MgCl2, 16 mM ATP, 10 mM DTT, 1 mM NaN3, pH 6.5 at 25˚C.


5. People involved with sample collection, processing, analysis and/or submission:
	Sample preparation, collection, processing and analysis were performed by Li GC and Muretta JM.
	Olivieri C organized the data and submitted to DRUM.

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DATA-SPECIFIC INFORMATION FOR: SteadyState_Fluo_PKA+PKI_FRET.xlsx
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1. Number of variables:2
	Time(x) and Fluorescence intensity (I) 

2. Number of rows: 1013 rows

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DATA-SPECIFIC INFORMATION FOR: TR-FRET_PKI-PKA.xlsx
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1. Number of variables:2
	Time(x) and Fluorescence intensity (I) 


2. Number of rows: 2001