The Internal Protein Sequencing Research Committee invites you to
participate in a collaborative study involving identification of a SDS-PAGE
separated protein sample. One of the primary goals of this study is to
provide our members with a mechanism to independently and anonymously
evaluate their ability to identify SDS-PAGE separated proteins. For those
ABRF laboratories that do not currently offer protein identification as a
service, this study should also provide an attractive introduction to this
technology. For those facilities interested in trying these procedures and
who do not have an established protocol, a representative procedure and
references will be included with the sample.
The protein samples for this study will be subjected to SDS-PAGE by the
Committee and then will be distributed in the form of a Coomassie
Blue-stained SDS gel slice. As with many samples purified by one
dimensional SDS PAGE, this sample may contain a contaminating protein at a
somewhat lower level. A control gel slice will also be provided that
should not contain any added protein. Participants will be asked to
identify the protein(s) via any approach they chose. Approaches that might
be utilized (either individually or in combination) could include enzymatic
cleavage followed by:
1. MALDI-MS and peptide mass searching
2. Preparative HPLC/Edman degradation of one or more purified peptides
3. Analytical HPLC/MS/MS followed by searching of MS/MS fragmentation ion
data
We anticipate distributing the samples in October, and request that the
resulting data be returned in December of 1997 so that sufficient time will
be available to tabulate and present the results at the 1998 ABRF Meeting
(March 21-24 in San Diego). Since preparation of these samples will
involve a reasonable amount of effort, the purpose of this notice and a
parallel mailing to ABRF Directors and Corporate sponsors is to identify
those member laboratories and sponsors that would like to receive these
samples and that intend to participate. The reason this sample will be
distributed only in the form of a stained gel slice is to try to keep the
Committee's workload reasonable, it does not imply in gel digestion is
generally any more nor less efficacious than on-PVDF digestion.
Please note that this sample will only be sent to those ABRF Directors and
Corporate Sponsors who request it via returning the postcard that is being
mailed this week or via sending an email message (marked ABRF Internal
Protein Sequence Sample) to Ms. Ann Ewing (ann.ewing@yale.edu) that is
received by October 24, 1997.
As with past ABRF studies, the resulting data will be returned via a third
party to protect the anonymity of the respondents and summaries of the
results of this study will be provided to all participants. In closing, we
thank you for your support of the ABRF and we encourage you to participate
in this unique study.
Sincerely,
The ABRF Internal Protein Sequencing Research Committee
Roland Annan, SmithKline Beecham
Karen DeJongh, Zymogenetics
Carol Fiol, Colorado State University
Ulf Hellman, Ludwig Institute
Scott Patterson, Amgen
Dave Speicher, Wistar Institute
Kristine Swiderek, Beckman Research Inst.
Ken Williams,Yale University