Molecular cloning and expression analysis of P-selectin glycoprotein ligand-1 from zebrafish (Danio rerio)

Fish Physiol Biochem. 2012 Apr;38(2):555-64. doi: 10.1007/s10695-011-9535-7. Epub 2011 Jul 14.

Abstract

To date, the best characterized glycoprotein ligand for P-selectin is P-selectin glycoprotein ligand-1 (PSGL-1). In this study, we cloned the full-length cDNA of PSGL-1 from zebrafish (Danio rerio). Zebrafish PSGl-1 cDNA is 1,594 bp and encodes a putative 284 amino acid protein with a theoretical molecular weight of 30.33 kDa and isoelectric point of 7.96. A signal peptide of 27 amino acids is predicted. The putative protein contains an extracellular mucin-like domain, a transmembrane domain and a cytoplasmic domain, with homology to mammalian PSGL-1. In the putative P-selectin binding region, there are 1 potential tyrosine sulfation site and 12 potential threonine O-glycosylation sites. A single extracellular cysteine, at the junction of the extracellular and transmembrane domains, suggests a disulfide-bonding pattern. The amino acid sequence of zebrafish PSGL-1 is 19-22% identical to that of mammalian PSGL-1. RT-PCR and whole-mount in situ hybridization analysis revealed that zebrafish PSGL-1 was expressed in early embryonic development, and the expression has an increased trend from 0.2 (1-cell stage) to 72 hpf. The results indicate that the general domain structure of PSGL-1 protein is conserved among species, and zebrafish PSGL-1 plays important roles in embryonic development and probably has similar biological function to that of mammalian PSGL-1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • Gene Expression
  • Membrane Glycoproteins / genetics*
  • Membrane Glycoproteins / metabolism
  • Molecular Sequence Data
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Zebrafish / embryology
  • Zebrafish / genetics*
  • Zebrafish / metabolism

Substances

  • Membrane Glycoproteins
  • P-selectin ligand protein