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Protein Sequencing Research Group (PSRG)

Mission

The mission of the Protein Sequencing Research Group (PSRG) is to design studies to assess techniques for determining the primary structure of protein termini that will allow participating laboratories to gauge their expectations and competence in performing these analyses. 

PSRG Guidelines:
The PSRG has the responsibility to regularly design and distribute samples to the ABRF membership following a reasonable time line decided mutually by the PSRG and the Executive Board of ABRF (EB). This includes sample design (EB approval required), sample preparation, test evaluation by PSRG members, sample distribution, data receipt and evaluation, oral and/or poster presentation at the ABRF or other international meeting (e.g., Protein Society or ASBMB), and finally, publication in a peer reviewed journal. The study should be designed for N – and/or C-terminal sequence analysis of proteins containing the 20 common amino acids plus (optionally) other post-translationally modified amino acids that are commonly found in proteins. Any technology that yields information about the termini of proteins may be used. The PSRG should be comprised of 6-8 members plus one EB Liaison whose purpose is to assist the research group and to facilitate communication with the EB. The PSRG should meet about once a month via a conference call to discuss the current sample and ensure continued progress. Once a year, one of the members will be chosen as chairperson in a fashion determined by the PSRG. The chair has the ultimate responsibility for ensuring the success of the RG and its studies, from initial design to the final publication. This includes communication with the EB, management office, JBT or other journal, chairs of the other protein research groups, ABRF membership, and the yearly ABRF meeting organizers. Every year, 2-3 members rotate off the RG and are replaced by new members who are nominated by the PSRG for approval by the EB. The PSRG should use this as a guideline but is not limited to it, either in order presented or type of sample. To help ensure that the PSRG meets the continuing needs of ABRF members, each sample submission form would contain a few survey questions to better gauge the kinds of samples that would be most useful to our members and to solicit suggestions for future studies.

Current Membership

Hediye Bromagehediyebromage@gmail.comChair
[Greg Cavey]gscavey@kazoosmic.com 
Robert Englishrdenglish@shimadzu.com 
Brian Feildbjfeild@shimadzu.com 
Xuemei Luoxueluo@utmb.edu 
Sara McgrathSara.Mcgrath@fda.hhs.gov 
David Wooddwood12@slu.edu 
Allis Chienallis.chien@my.abrf.orgEB Liaison

Studies

1)PSRG 2014-2015 Study: N-terminal sequencing of standard proteins by dimethyl labeling and bottom-up mass spectrometry
    - View Study Announcement (34K)
    - View Sample Insert (359K)
2)PSRG 2013-2014 Study: N-terminal sequencing of standard proteins by N-terminal labeling and mass spectrometry
    - View Study Announcement (30K)
    - View Sample Insert (237K)
3)PSRG 2012-2013 Study : Sample preparation and terminal sequencing of a protein mixture
    - View Document 2012-2013 Study Announcement 
    - View Document 2012 Participant Results Survey 
4)PSRG 2011 Study: Comparing sensitivity limits for N-terminal Sequencing Technologies
    - View PSRG 2011 Study Announcement 
    - View PSRG 2011 Study Sample Insert (36K)
5)PSRG 2010 Study: Comparing Edman and MS Sequencing of a Monoclonal Antibody (study closed)
    - View Study Announcement (94K)
    - View Cover Letter/Instructions (20K)
    - View Google Doc's data reporting form and survey 
6)ESRG 2009 Study: Comparing Edman and Mass Spectrometry Techniques for N-terminal Sequencing
    - ESRG 2009 Study Announcement 
    - Cover Letter/Sample Instructions (15K)
    - Vendor ABRF Study Participation Guidelines (18K)
    - Data Sheet - Excel file for both Edman and MS Data 
7)ESRG 2008 Study "Investigation into homopolymeric amino acid N-terminal sequence tags and their effects on automated Edman degradation"
    - ESRG 2008 Study Announcement 
    - Cover Letter/Sample Instructions 
    - Data Sheet (41K)
    - Vendor ABRF Study Participation Guidelines (18K)
8)ESRG 2007 Study "Procedures used to Determine the N-Terminal Sequence of a Blocked Protein"
    - ESRG 2007 Study Announcement 
    - Cover Letter/Sample Instructions 
    - Deblocking Cycle 
    - Data Sheet (25K)
9)ESRG 2006 Study "Edman Sequencing as a Method for Polypeptide Quantitation"
    - ESRG 2006 Study Announcement 
10)ESRG 2005 Study "Identification of Modified Amino Acids by Edman Sequencing"
    - Instructions for Filling Out the Datasheet 
    - Sample Description Including Modified Amino Acids 
    - Data Sheet (19K)
11)ESRG 2004 Study "Modified Amino Acids in Edman Sequencing"
    - Sample Description Including Modified Amino Acids 
12)ESRG 2004 Study Aid: Pages 10-12 from "Identification of Modified PTH-Amino Acids in Protein Sequence Analysis" 1st ed. compiled by Mark Crankshaw and Greg Grant
    - Chromatogram: page 10 
    - Legend to chromatogram: page 11 
    - Legend continued: page 12 

Activities

1)ESRG Newsletter Vol.1 No.1, - Report on the ESRG at ABRF 2008, Salt Lake City, Utah and a Tribute to Pehr Edman

Electronic Posters

 PSRG 2015 - ABRF Poster and Oral Presentations: N-terminal Identification of a Standard Protein at Low Picomole Levels by Dimethyl Labeling and Bottom-up Mass Spectrometry
 PSRG 2014 - ABRF Poster and Oral Presentations: N-terminal Sequencing of Standard Proteins by N-terminal Labeling and Mass Spectrometry
 PSRG 2013 - ABRF Poster and Oral Presentations: Results of the PSRG 2013 Study Year 2: Terminal Sequencing of Standard Proteins in a Mixture
 PSRG 2012 - ABRF Poster and Oral Presentations: Results of the PSRG 2012 Study: Terminal Sequencing of Standard Proteins in a Mixture, Year 1 of the 2 Year Study
 PSRG 2011 - ABRF Presntations: Results of the PSRG 2011 Study: SensitivityAssessment of Edman and Mass Spectrometric Terminal Sequencing of an Undisclosed Protein
    - PSRG 2011 Poster (4,387K)
    - PSRG 2011 Oral Presentation (9,213K)
 PSRG 2010 Poster: Edman and Mass Spectrometric Terminal Sequencing of a Monoclonal Antibody
 PSRG 2010 Oral Presentation: N-Terminal Sequencing of an Antibody
 ESRG 2009 Poster: Comparison of Edman and Mass Spectrometry Techniques for N-terminal Sequencing
 ESRG 2009 Oral Presentation. "N-Terminal Sequencing Techniques"
 ESRG 2008 Oral Presentation. "Effects of a Homopolymeric Amino Acid Tag on Edman Degradation"
 ESRG 2008 Poster. "Investigation into Homopolymeric Amino Acid N-terminal Sequence Tags and their Effects on Automated Edman Degradation"
 ESRG 2007 Poster: "Procedures used to Determine the N-Terminal Sequence of a Blocked Protein"
 ESRG 2007 Oral Presentation: "Procedures used to Determine the N-Terminal Sequence of a Blocked Protein"
 ESRG 2006 poster: "Edman Sequencing as a Method for Polypeptide Quantitation"
 ESRG 2006: Oral presentation: Edman Sequencing as a Method for Polypeptide Quantitation
 ESRG 2005 "State of Edman Sequencing" Survey, used for both the poster and oral presentation at the ABRF meeting, Savannah, GA, February 5-8, 2005
 ESRG 2005 poster "Identification of Modified Amino Acids by Edman Sequencing"
 ESRG 2005: Oral presentation: Modified amino acids by Edman sequencing
 "ABRF ESRG 2004 Study: Modified Amino Acids in Edman Sequencing"; ESRG study poster presented at the 2004 ABRF meeting February 28-March 2 in Portland, Oregon.
 "Modified Amino Acids in Edman Sequencing"; ESRG study results presented at the 2004 ABRF meeting February 28-March 2 in Portland, Oregon.
 "ABRF-ESRG 2003: Analysis of a PVDF-Bound Known Protein with a Homogeneous Amino-Terminus" ; ESRG study poster presented at the 2003 ABRF meeting February 10-13 in Denver, Colorado.
 Slides from tutorial, "Making the Most of Your Edman Sequence Data; A Primer on Data Calling, Analysis, Interpretation and Reporting" in pdf format, presented at the 2003 ABRF meeting February 10-13 in Denver, Colorado.
 Slides from the 2003 ESRG Study oral presentation given at the 2003 ABRF meeting, Feb 10-13 in Denver, Colorado.
 "ABRF-2002 ESRG, a Difficult Sequence: Analysis of a PVDF-Bound Known Protein With a Heterogeneous Amino-Terminus"; poster presented at the 2002 ABRF meeting March 10-12 in Austin, Texas.
 Slides from ESRG 2002 Study oral presentation given at 2002 ABRF meeting March 10-12 in Austin, Texas
 The first half of the tutorial session "N-terminal Edman sequencing sample preparation" slides in pdf format.
 Second half of the tutorial session "N-terminal Edman sequencing sample preparation" slides in pdf format.
 Here is the method briefs handout from the tutorial session. Please use these methods and references only as guides for you to develop similar methods more fully in your own labs, no liability to ABRF for their use.

Membership History

Member NameOrganizationDetails
Daniel BruneArizona State Univ Chair: 02/05 - 02/06
Member: 02/06 - 04/07
Scott BuckelAmgen, Inc. Member: 01/01 - 03/03
Richard G. CookBaylor Col of Med Member: 01/01 - 03/04
J. M. CrawfordYale Univ. Keck Biotech Res. Lab. Member: 01/01 - 03/04
Dr Nancy D DenslowUniv of Florida-Gainesville Chair: 03/04 - 02/05
Member: 03/02 - 03/04
EB liaison: 02/05 - 02/09
David R. DupontApplied Biosystems Member: 01/01 - 03/02
Dr Robert EnglishUniv. of Texas Medical Branch Chair_(co-chair): 04/12 - 04/13
Member: 05/11 - 04/12
Dr Hediye Erdjument-BromageSloan Kettering Inst Member: 04/14 - 04/15
Joseph FernandezRockefeller Univ Chair: 01/01 - 03/02
Member: 03/02 - 03/03
Mr. Mark K. GarfieldNIAID/NIH Member: 05/12 - 04/15
Brian HamptonUniverstiy of Maryland School of Medicine Member: 02/05 - 04/07
Member: 02/08 - 02/09
Ad hoc: 02/09 - 04/10
Co-Chair: 04/07 - 02/08
Dr William G HendricksonUniv. of Illinois, Chicago EB Liason: 03/13 - 04/14
Peter HunzikerUniversity of Zurich Chair: 02/08 - 02/09
MemberBecome Ad Hoc: 02/09 - 04/10
Member: 03/06 - 02/08
Ad hoc: 04/10 - 04/11
Pegah R JaliliSigma-Aldrich Member: 06/11 - 04/14
Viswanatham KattaGenentech Member: 06/10 - 03/13
Ryuji KobayashiUT MD Anderson Cancer Center Member: 03/03 - 03/06
William S. LaneHarvard University EB Liason: 10/03 - 02/05
Joseph W. LeonePfizer Chair: 02/06 - 04/07
Member: 03/04 - 02/06
Member chair-emeritus: 04/07 - 02/08
Joseph F. LeykamMichigan State University Member: 04/07 - 08/07
Dr. Klaus D. LinseBiosynthesis Inc. Member: 02/05 - 02/08
Benjamin J MaddenMayo Clinic Chair: 02/03 - 03/04
Member: 03/04 - 02/05
Member: 01/01 - 02/03
Kwasi G MawuenyegaWashington University School of Medicine Member: 05/09 - 04/12
Dr. Sara C. McGrathUS FDA Chair_(co-chair): 04/13 - 04/15
Member: 05/12 - 03/13
Dr. Ejvind MortzAlphalyse : 03/13 - 04/14
John M NeveuHarvard University Chair: 04/02 - 01/03
Member: 01/01 - 03/02
Member: 02/03 - 02/05
Dr. Ronald L. NieceResearch Resources & Technologies Member: 03/88 - 03/91
Dr. Brett S PhinneyProteomics Core UC Davis Genome Center EB Liason: 04/14 - 04/15
Dr Jan PohlEmory Univ Sch of Med Member: 03/04 - 04/07
Dr. Henriette A. RemmerUniversity of Michigan Member: 06/10 - 04/11
Ad hoc: 03/13 - 04/15
co-chair: 04/11 - 04/12
Co-chair: 04/12 - 03/13
Wendy SandovalGenentech, Inc. Member: 04/10 - 04/11
Member changing to cochair: 04/07 - 02/09
Co-chair: 02/09 - 04/10
Jack SimpsonUS Pharmacopeia EB liaison: 02/09 - 03/13
John S. SmithUniv of Texas Med Branch Member: 04/10 - 04/11
Member changing to cochair: 09/07 - 02/09
Co-chair: 02/09 - 04/10
Laurey SteinkeUniversity of Nebraska Medical Center EB liason: 01/01 - 09/03
Dr. Detlev SuckauBruker Daltonik GmbH MemberBruker Daltonics: 05/10 - 03/13
Richard S ThomaMonsanto Member: 02/08 - 02/09
Member: 02/05 - 04/07
Ad hoc: 02/09 - 04/10
Co-Chair: 04/07 - 02/08
James WaltersSigma-Aldrich Chair: 04/10 - 04/11
Member: 04/09 - 04/10
Ad hoc: 04/12 - 03/13
co-chair: 04/11 - 04/12
Ken R. WilliamsYale Univ, Keck Lab Member: 03/88 - 03/90
Dr. Bosong XiangMonsanto Company Member: 04/09 - 04/12

Publications

  1. A Synthetic Peptide for Evaluating Protein Sequencing Capabilities: Design of ABRF-93SEQ and Results. In (J.W. Crabb, ed.) Techniques in Protein Chemistry V, Academic Press, San Diego, pp133-141 (1994)
    Rush, J., Andrews, P. C., Crimmins, D. L., Gambee, J. E., Grant, G. A., Mische, S. M. and Speicher, D. W.
  2. ABRF ESRG 2005 Study: identification of seven modified amino acids by Edman sequencing. J Biomol Tech, 2006. 17(5): p. 308-326.
    Brune, D., N.D. Denslow, R. Kobayashi, W.S. Lane, J.W. Leone, B.J. Madden, J.M. Neveu, and J. Pohl
  3. ABRF-2002 ESRG, A Difficult Sequence: Analysis of a PVDF-Bound KNown Protein With a Heterogeneous Amino-Terminus Journal of Biomolecular Techniques 13:246-257 (2002).
    Buckel, S.D., Cook,R.G., Crawford, J.M., Dupont, D.R., Madden,B.J., Neveu,J.M., Steinke, L., Fernandez, J
  4. ABRF-2003 ESRG: Analysis of a PVDF-Bound KNown Protein With a Heterogeneous Amino-Terminus Journal of Biomolecular Techniques 14:278-288 (2003).
    Buckel, S.D., Cook,R.G., Crawford,M.J., Denslow, N., Madden,B.J., Steinke, L., Fernandez, J., Neveu,J.M
  5. ABRF-97SEQ: Sequencing Results of a Low-Level Sample. J Biomol Tech, 10:26-32 (1999)
    Stone, K., Fernandez, J., Admon, A., Henzel, W., Lane, W., Rohde, M., and Steinke, L.
  6. ABRF-98SEQ: Evaluation of peptide sequencing at high sensitivity J. Biomol. Tech., Jun 2000; 11: 92 - 99.
    WJ Henzel, A Admon, SA Carr, G Davis, K De Jongh, W Lane, M Rohde, and L Steinke
  7. Amino Acid Analysis and Sequencing - What Is State-of-the-Art?
    Niece, R.L., L.H. Ericsson, A.V. Fowler, A.J. Smith, D.W. Speicher, J.W. Crabb, and K.R. Williams.

    "Methods in Protein Sequence Analysis" (H. Jörnvall, J.-O. Höög, and A.-M. Gustavsson, Eds.), Birkhäuser Verlag, Basel, 133-141, 1991

  8. Assignment of Cysteine and Tryptophan Residues during Protein Sequencing: Results of ABRF-94SEQ. In (J. W. Crabb, ed.) Techniques in Protein Chemistry VI, Academic Press, San Diego, pp 209-217 (1995)
    Gambee, J. E., Andrews, P. C., Grant, G. A., Merrill, B., Mische, S. M. and Rush, J.
  9. Design and Analysis of ABRF-95SEQ, a Recombinant Protein with Sequence Heterogeneity. In (D. Marshak, ed.) Techniques in Protein Chemistry VII, Academic Press, San Diego, pp 347-358 (1996)
    DeJongh, K. S., Fernandez, J., Gambee, J. E., Grant, G. A., Merrill, B., Stone, K. L. and Rush, J.
  10. Design, Characterization and Results of ABRF-89SEQ: A Test Sample for Evaluating Protein Sequencer Performance in Protein Microchemistry Core Facilities. In (T. E. Hugli, ed.) Current Research in Protein Chemistry, Academic Press, pp 159-166 (1989)
    Speicher, D. W., Grant, G. A., Niece, R. L., Blacher, R. W., Fowler A. V. and Williams, K. R.
  11. Edman Sequencing Research Group 2004 Study: Modified Amino Acids in Edman Sequencing, J. Biomol. Tech., Sep 2005; 16: 272 - 284.
    D. Brune, J.M. Crawford, R.G. Cook, N.D. Denslow, R. Kobayashi, B.J. Madden, J. M. Neveu, and L. Steinke
  12. Evaluation of ABRF-96SEQ: A Sequence Assignment Exercise. In (D. R. Marshak, ed.) Techniques in Protein Chemistry VIII, Academic Press, San Diego, pp 69-78 (1997)
    Fernandez, J., Admon, A., DeJongh, K., Grant, G., Henzel, W., Lane, W. S., Stone, K. L. and Merrill, B.
  13. Evaluation of Protein Sequencing Core Facilities: Design, Characterization, and Results form a Test Sample (ABRF-91SEQ). In (R. H. Angeletti, ed.) Techniques in Protein Chemistry III, Academic Press, San Diego, pp 35-35 (1992)
    Crimmins, D. L., Grant, G. A., Mende-Muller, L. M., Niece, R. L., Slaughter, C., Speicher, D. W. and Yuksel, K. U.
  14. Instrumentation Used in Protein and Nucleic Acid Resource Facilities: A Survey of Users
    Beach, C.M. G.M. Hathaway, T.K. Hayes, A.J. Smith, and R.L. Niece.

    ABRF News, Vol. 2.1 Supplement: 1-7, 1991

  15. Protein Sequencing from Polyvinylidene Difluoride Membranes: Design and Characterization of a Test Sample (ABRF-90SEQ) and Evaluation of Results. In (J. J. Villafranca, ed.) Techniques in Protein Chemistry II, Academic Press, San Diego, pp 151-162 (1991)
    Yuksel, K. U., Grant, G. A., Mende-Muller, L. M., Niece, R. L., Williams, K. R. and Speicher, D. W.
  16. Protein Sequencing of Post-translationally Modified Peptides and Proteins: Design, Characterization and Results of ABRF-92SEQ. In (R. H. Angeletti, ed.) Techniques in Protein Chemistry IV, Academic Press, San Diego, pp 453-461 (1993)
    Mische, S. M., Yuksel, K. U., Mende-Muller, L. M., Matsudaira, P., Crimmins, D. L. and Andrews, P. C.
  17. Synthetic Peptide for Evaluating Protein Sequencer and Amino Acid Analyzer Performance in Core Facilities: Design and Results
    Niece, R.L., K.R. Williams, C.L. Wadsworth, J. Elliott, K.L. Stone, W.J. McMurray, A. Fowler, D. Atherton, R. Kutny, and A.J. Smith. A.

    in "Techniques in Protein Chemistry" (Hugli, T., Ed.), Academic Press, San Diego, 89-110, 1989

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