Proteome of the large Pseudomonas myovirus 201φ2-1: Delineation of proteolytically processed virion proteins

Julie A. Thomas, Susan T. Weintraub, Kevin Hakala, Philip Serwer, Stephen C. Hardies

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

Pseudomonas chlororaphis phage 201φ2-1 produces a large structurally complex virion, including the products of 89 phage genes. Many of these proteins are modified by proteolysis during virion maturation. To delineate the proteolytic maturation process, 46 slices from an SDS-polyacrylamide gel were subjected to tryptic digestion and then HPLC-electrospray ionization-tandem mass spectrometry analysis. The scale of the experiment allowed high sequence coverage and detection of mass spectra assigned to peptides with one end produced by trypsin and the other end derived from a maturation cleavage (semitryptic peptides). Nineteen cleavage sites were detected in this way. From these sites, a cleavage motif was defined and used to predict the remaining cleavages required to explain the gel mobility of the processed polypeptide species. Profiling the gel with spectrum counts for specific polypeptide regions was found to be helpful in deducing the patterns of proteolysis. A total of 29 cleaved polypeptides derived from 19 gene products were thus detected in the mature 201φ2-1 virion. When combined with bioinformatics analyses, these results revealed the presence of head protein-encoding gene modules. Most of the propeptides that were removed from the virion after processing were acidic, whereas the mature domain remaining in the virion was nearly charge-neutral. For four of these processed virion proteins, the portions remaining in the mature virion were mutually homologous. Spectrum counts were found to overestimate the relative quantity of minor polypeptide species in the virion. The resulting sensitivity for minor species made it possible to observe a small amount of general proteolysis that also affected the virions.

Original languageEnglish (US)
Pages (from-to)940-951
Number of pages12
JournalMolecular and Cellular Proteomics
Volume9
Issue number5
DOIs
StatePublished - May 2010

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Molecular Biology

Fingerprint Dive into the research topics of 'Proteome of the large Pseudomonas myovirus 201φ2-1: Delineation of proteolytically processed virion proteins'. Together they form a unique fingerprint.

Cite this