RT-PCR- and MALDI-TOF mass spectrometry-based identification and discrimination of isoforms homologous to pufferfish saxitoxin- and tetrodotoxin-binding protein in the plasma of non-toxic cultured pufferfish (Takifugu rubripes)

Biosci Biotechnol Biochem. 2013;77(1):208-12. doi: 10.1271/bbb.120701.

Abstract

Four genes of Takifugu rubripes, tentatively designated Tr1-Tr4, encoding homologs of pufferfish saxitoxin- and tetrodotoxin-binding protein, were identified by BLAST search and 3'-RACE. RT-PCR and MALDI-TOF mass spectrometry allowed the identification and discrimination of Tr isoforms from the non-toxically cultured specimens. The expression of Tr1 and Tr3 mRNAs exclusively in the liver and the presence of their products as 120-kDa plasma proteins were confirmed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Blood Proteins / chemistry*
  • Blood Proteins / genetics
  • Blood Proteins / metabolism
  • Fish Proteins / chemistry*
  • Fish Proteins / genetics
  • Fish Proteins / metabolism
  • Liver / chemistry
  • Liver / metabolism*
  • Molecular Sequence Data
  • Protein Binding
  • Protein Isoforms / chemistry
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Saxitoxin / metabolism
  • Sequence Alignment
  • Sodium Channels / chemistry*
  • Sodium Channels / genetics
  • Sodium Channels / metabolism
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Takifugu / blood*
  • Tetrodotoxin / metabolism

Substances

  • Blood Proteins
  • Fish Proteins
  • Protein Isoforms
  • Sodium Channels
  • tetrodotoxin-binding protein
  • Saxitoxin
  • Tetrodotoxin