Proteomic analysis of the neutrophil proteins of the tammar wallaby (Macropus eugenii)

Kiran Ambatipudi, Julie Old, Michael Guilhaus, Mark Raftery, Lyn Hinds, Elizabeth Deane*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

Abstract

A proteomic analysis of neutrophils from the tammar wallaby, Macropus eugenii, has been performed. Neutrophils were isolated from peripheral blood using density gradient centrifugation with Histopaque-1077, followed by treatment with ammonium chloride to lyse residual erythrocytes. Two-dimensional gel electrophoresis (2-DE) of lysed neutrophils was undertaken followed by in-gel trypsin digest and nanoliquid chromatography coupled tandem mass spectrometry (LC-MS) analysis and database searches. Seventy-seven proteins were isolated, 53 of which could be identified with high confidence as primarily of cytosolic origin. Protein identifications were only possible by matching identical peptide sequences within the NCBInr mammalian database with the Mascot search program. Sequence identities were only deemed acceptable if more than three peptides were identified, the precursor/protein ion tolerances were less than ± 0.25 Da and the total Mowse scores were greater than 100. The validity of this approach was tested using a scrambled database where no single identified peptide showed Mowse scores greater than 55. This is the first report of the neutrophil proteins of any marsupial and represents a first step in examining the identity of proteins involved in innate defence in this marsupial.

Original languageEnglish
Pages (from-to)283-291
Number of pages9
JournalComparative Biochemistry and Physiology - Part D: Genomics and Proteomics
Volume1
Issue number3
DOIs
Publication statusPublished - Sep 2006

Keywords

  • Electrospray ionisation-tandem mass spectrometry
  • Marsupials
  • Neutrophils
  • Two-dimensional gel electrophoresis

Fingerprint

Dive into the research topics of 'Proteomic analysis of the neutrophil proteins of the tammar wallaby (Macropus eugenii)'. Together they form a unique fingerprint.

Cite this