Over 500 people attended the sixth ABRF Satellite Meeting held on June 22, 1991 in conjunction with the annual symposium of the Protein Society in Baltimore. ABRF gratefully acknowledges the efforts of Beth Fowler who organized the satellite meeting as well as the continuing support of the Protein Society and especially Kristen Copley in coordinating the satellite meeting and required facilities.
Each ABRF research committee presented an oral summary of their 1991 studies. In addition, poster presentations by the committees featured more detailed data and provided opportunities for extensive discussion during the refreshment break as well as after the formal presentations.
The peptide synthesis/mass spectrometry subcommittee, in their first year of operation, conducted an ambitious study in which they asked core facilities to synthesize and submit the peptide V-K-K-R-C-S-M-W-I-I-P-T-D-D-E-A for detailed characterization. The committee members then subjected the submitted samples to the following analyses: amino acid analysis, analytical HPLC, capillary electrophoresis. electrospray and plasma desorption mass spectrometry, and the ability to serve as a substrate for casein kinase II and cyclic AMP-dependent protein kinases. Thirty-six laboratories submitted samples for analysis; this is nearly 40% of the ABRF member laboratories that indicated they perform peptide synthesis. The results showed that the standard peptide could be synthesized successfully using either the tBOC or Fmoc approach; however, the ABRF core laboratories that participated in the study succeeded more frequently with the Fmoc approach.
The study suggested that a quantitative separation method coupled with a mass spectrometric technique provides the best approach for determining the purity and the authenticity of the synthesized product. This is the first time that a study of peptide synthesis capabilities has been conducted of such a large magnitude. The substantial contributions of both the committee members and the large number of participating member laboratories have contributed to an important, innovative, and unique study. The results were presented by Steve Carr, Al Smith, Lynn Williams, and Janis Young.
The survey subcommittee report, presented by Carol Beach, summarized the results of the 1991 survey on training and education. The information will be helpful in suggesting what types of education activities ABRF might undertake in the future.
The Amino Acid Analysis subcommittee reported results for ABRF-9lAAA. This year's sample, bovine serum albumin, was distributed in sufficient quantity to allow members to determine the best possible composition and to perform cysteine and tryptophan determinations. The best compositions compared favorably with the known composition. However, cysteine, tryptophan, and methionine values were less accurate than for the other 15 amino acids, indicating that there is room for improvement in the determination of these residues. This data was presented by Ray Paxton, Dan Strydom, and George Tarr. The efforts of Sam Margolis from the National Institute of Standards and Technology who worked closely with the committee in developing this study are gratefully appreciated.
Our guest speaker, I)r Gregory Buck of Virginia Commonwealth University, described key factors in organizing and running a successful nucleic acids core facility which provides oligonucleotide synthesis and DNA sequencing services to the research community.
Following the lectures, David Speicher chaired an open discussion of ABRF activities and future directions. Several important issues were raised. There was significant interest from the audience for the purchase of additional aliquots of amino acid and sequence standards. Sam Margolis reported that this year's amino acid standard can be purchased directly from NIST. In future years, the committees will prepare a sufficiently large number of samples for purchase after the initial study, if this is feasible. There was also substantial sentiment that a standard notation is needed for reporting sequences and especially designating confidence levels. A related concern was the low average accuracy of positive sequence calls in the 1991 study (87% accurate). Another concern was that laboratories need more direct feedback on where their data ranks in the study. The peptide synthesis/mass spectrometry and the amino acid analysis committees plan to return results directly to individual laboratories.
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