A novel dimer of a C-type lectin-like heterodimer from the venom of Calloselasma rhodostoma (Malayan pit viper)

Runhua Wang, Chunguang Kong, Prasanna Kolatkar, Maxey C M Chung

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10 Citations (Scopus)

Abstract

We have isolated a potent platelet aggregation inducer from the crude venom of Calloselasma rhodostoma (Malayan pit viper), termed rhodoaggretin, with a novel oligomeric structure consisting of a dimer of C-type lectin-like heterodimers. On the basis of its native molecular mass of 66 kDa, and a Mr of 30 kDa for its disulfide-linked αβ-heterodimer, we propose that rhodoaggretin exists as a (αβ)2 complex in the native state. We postulate that the di-dimer is stabilized by non-covalent interactions as well as by an intersubunit disulfide bridge between the two αβ-heterodimers. This conclusion is based on the following observations: (a) sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the non-reduced rhodoaggretin gave a major 28 and a minor 52 kDa band. (b) Prior treatment of rhodoaggretin with a limited amount of 2-mercaptoethanol (2-ME; 0.1%) resulted in the complete abolishment of the 52 kDa band in SDS-PAGE. (c) Two-dimensional SDS-PAGE in the presence of 3% 2-ME showed that both the 28 and 52 kDa bands gave two bands each with Mrs of 18 (α-subunit) and 15 (β-subunit) kDa. (d) Mass spectrometric analyses showed that purified rhodoaggretin had a Mr of 30 155.39±3.25 Da while its s-pyridylethylated α- and β-subunits had Mrs of 16 535.62±2.98 and 15 209.89±1.61 Da respectively. These molecular weight data suggested the presence of 15 cysteinyl residues in rhodoaggretin as compared to the 14 that are reported for the heterodimeric C-type lectin-like proteins. This extra cysteinyl residue is a candidate for the formation of the intersubunit disulfide bond in the (αβ)2 complex. (e) Homology structural modeling studies showed that the extra cysteinyl residue can indeed form a disulfide bond that covalently links the two αβ-heterodimers as proposed above.

Original languageEnglish
Pages (from-to)447-453
Number of pages7
JournalFEBS Letters
Volume508
Issue number3
DOIs
Publication statusPublished - 23 Nov 2001
Externally publishedYes

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Keywords

  • C-type lectin-like protein
  • Calloselasma rhodostoma
  • Molecular modeling
  • Platelet aggregation
  • Rhodoaggretin
  • Sequence homology

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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