Characterization and evolution of a gene encoding a Trimeresurus flavoviridis serum protein that inhibits basic phospholipase A2 isozymes in the snake's venom

Ikuo Nobuhisa, Satoshi Inamasu, Makoto Nakai, Akira Tatsui, Tomohiro Mimori, Tomohisa Ogawa, Yasuyuki Shimohigashi, Yasuyuki Fukumaki, Shosaku Hattori, Hiroshi Kihara, Motonori Ohno

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

The proteins that bind phospholipase A2 (PLA2) isozymes of Trimeresurus flavoviridis (habu snake, crotalinae) venom were fractionated from sera on four columns, each conjugated with one of four PLA2 isozymes. Five proteins, termed PLA2 inhibitors (PLI) I-V, were obtained as the binding components. The combinations of the binding components differed depending on the PLA2 isozymes. PLI-IV and PLI-V correspond to PLI-A and PLI-B, respectively, which were known to bind to a major [Asp49]PLA2, PLA2, and contained a segment similar to the carbohydrate-recognition domain of C-type lectins. PLI-I, which is a major component of inhibitory proteins against three basic PLA2 isozymes, PLA-B (a basic [Asp49]PLA2) and basic proteins I and II (both [Lys49]PLA2s), has been isolated, and its partial amino acid sequence has been determined. A cDNA encoding PLI-I was isolated from a T. flavoviridis liver cDNA library and sequenced. PLI-I cDNA encoded 200 amino acid residues, including a signal peptide of 19 amino acid residues. One sugar chain was predicted to occur at position 157. A gene coding for PLI-I was isolated. It is 9.6-kb long and consists of five exons and four introns. Comparison of the exon-intron structure of the PLI-I gene with those of genes encoding urokinase-type-plasminogen-activator receptor (uPAR), Ly-6, CD59 and neurotoxins showed that they have characteristic unit encoding approximately 90 amino acid residues, which is divided over two exons. This strongly suggests that the PLI-I gene belongs to the uPAR, Ly-6, CD59 and neurotoxin gene family. There are two types of structurally different inhibitors against PLA2 isozymes in T. flavoviridis serum with different evolutionary origins.

Original languageEnglish
Pages (from-to)838-845
Number of pages8
JournalEuropean Journal of Biochemistry
Volume249
Issue number3
DOIs
Publication statusPublished - 1997 Jan 1
Externally publishedYes

Keywords

  • Affinity column
  • Amino acid sequence
  • Crotalinae snake
  • Liver
  • cDNA

ASJC Scopus subject areas

  • Biochemistry

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