Affordable Access

deepdyve-link
Publisher Website

Characterization of the mature form of a β-defensin-like peptide, Hoa-D1, in the lobster Homarus americanus.

Authors
  • Vu, Giap H1
  • Do, Daniel1
  • Rivera, Cindy D1
  • Dickinson, Patsy S2
  • Christie, Andrew E3
  • Stemmler, Elizabeth A4
  • 1 Department of Chemistry, Bowdoin College, 6600 College Station, Brunswick, ME 04011, United States. , (United States)
  • 2 Department of Biology, Bowdoin College, 6500 College Station, Brunswick, ME 04011, United States. , (United States)
  • 3 Békésy Laboratory of Neurobiology, Pacific Biosciences Research Center, School of Ocean and Earth Science and Technology, University of Hawaii at Manoa, 1993 East-West Road, Honolulu, HI 96822, United States. , (United States)
  • 4 Department of Chemistry, Bowdoin College, 6600 College Station, Brunswick, ME 04011, United States. Electronic address: [email protected] , (United States)
Type
Published Article
Journal
Molecular immunology
Publication Date
Sep 01, 2018
Volume
101
Pages
329–343
Identifiers
DOI: 10.1016/j.molimm.2018.07.004
PMID: 30036799
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

We report on the characterization of the native form of an American lobster, Homarus americanus, β-defensin-like putative antimicrobial peptide, H. americanus defensin 1 (Hoa-D1), sequenced employing top-down and bottom-up peptidomic strategies using a sensitive, chip-based nanoLC-QTOF-MS/MS instrument. The sequence of Hoa-D1 was determined by mass spectrometry; it was found to contain three disulfide bonds and an amidated C-terminus. The sequence was further validated by searching publicly-accessible H. americanus expressed sequence tag (EST) and transcriptome shotgun assembly (TSA) datasets. Hoa-D1, SYVRScSSNGGDcVYRcYGNIINGAcSGSRVccRSGGGYamide (with c representing a cysteine participating in a disulfide bond), was shown to be related to β-defensin-like peptides previously reported from Panulirus japonicas and Panulirus argus. We found Hoa-D1 in H. americanus hemolymph, hemocytes, the supraoesophageal ganglion (brain), eyestalk ganglia, and pericardial organ extracts, as well as in the plasma of some hemolymph samples. Using discontinuous density gradient separations, we fractionatated hemocytes and localized Hoa-D1 to hemocyte sub-populations. While Hoa-D1 was detected in semigranulocytes and granulocytes using conventional proteomic strategies for analysis, the direct analysis of cell lysates exposed evidence of Hoa-D1 processing, including truncation of the C-terminal tyrosine residue, in the granulocytes, but not semigranulocytes. These measurements demonstrate the insights regarding post-translational modifications and peptide processing that can be revealed through the MS analysis of intact peptides. The identification of Hoa-D1 as a widely-distributed peptide in the lobster suggests the possibility that it may be pleiotropic, with functions in addition to its proposed role as an antimicrobial molecule in the innate immune system. Copyright © 2018 Elsevier Ltd. All rights reserved.

Report this publication

Statistics

Seen <100 times