Purification, characterization, and cDNA cloning of a novel soluble saxitoxin and tetrodotoxin binding protein from plasma of the puffer fish, Fugu pardalis

Mari Yotsu-Yamashita, Atsuko Sugimoto, Takahiro Terakawa, Yuki Shoji, Teruo Miyazawa, Takeshi Yasumoto

Research output: Contribution to journalArticlepeer-review

86 Citations (Scopus)

Abstract

Some species of puffer fish have been reported to possess both of tetrodotoxin and saxitoxin, which share one binding site on sodium channels. We purified a novel soluble glycoprotein that binds to these toxins from plasma of the puffer fish, Fugu pardalis, and named puffer fish saxitoxin and tetrodotoxin binding protein (PSTBP). PSTBP possessed a binding capacity of 10.6 ± 0.97 nmol·mg-1 protein and a Kd of 14.6 ± 0.33 nM for [3H]saxitoxin in equilibrium binding assays. [3H]Saxitoxin (10 nM) binding to PSTBPs was half-inhibited by the presence of tetrodotoxin and saxitoxin at 12 μM and 8.5 nM, respectively. From the results of gel filtration chromatography (200 kDa) and SDS/PAGE (104 kDa), PSTBP was suggested to consist of noncovalently linked dimers of a single subunit. PSTBP was completely deglycosylated by glycopeptidase F, producing a single band at 42 kDa. Two highly homologous cDNAs to each other coding PSTBP (PSTBP1 and PSTBP2, the predicted amino-acid identity 93%), were obtained from a cDNA library of F. pardalis liver. These proteins consisted to two tandemly repeated homologous domains. The predicted amino-acid sequences of PSTBP1 and 2 were not homologous to that of saxiphilin, a reported saxitoxin binding protein, or sodium channels, but their N-terminus sequences were homologous to that of the reported tetrodotoxin binding protein from plasma of Fugu niphobles, which has not been fully characterized. The partially homologous cDNA sequences to PSTBP1 and 2 were also found in expressed sequence tag clones of nontoxic flounders liver. Presumably, PSTBP is involved in accumulation and/or excretion of toxins in puffer fish.

Original languageEnglish
Pages (from-to)5937-5946
Number of pages10
JournalEuropean Journal of Biochemistry
Volume268
Issue number22
DOIs
Publication statusPublished - 2001

Keywords

  • Puffer fish
  • Saxiphilin
  • Saxitoxin
  • Tetrodotoxin

ASJC Scopus subject areas

  • Biochemistry

Fingerprint Dive into the research topics of 'Purification, characterization, and cDNA cloning of a novel soluble saxitoxin and tetrodotoxin binding protein from plasma of the puffer fish, Fugu pardalis'. Together they form a unique fingerprint.

Cite this