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Fisheries Science

, Volume 75, Issue 3, pp 743–753 | Cite as

Cloning, expression, and characterization of a novel anti-HIV lectin from the cultured cyanobacterium, Oscillatoria agardhii

  • Tomomi Sato
  • Kanji Hori
Original Article Chemistry and Biochemistry

Abstract

OAA, the potent anti-HIV protein from Oscillatoria agardhii NIES-204 belongs to a new lectin family, shows strict binding specificity for high-mannose N-glycans, and has an extremely high association constant in the picomolar range for recombinant gp120, an envelope protein of HIV. In this study we have cloned the gene encoding OAA from the genomic DNA of the cyanobacterium, and efficiently expressed the recombinant lectin (rOAA) in Escherichia coli. The rOAA expressed as a His-tagged fusion protein was recovered in a soluble form and purified in high yield (48 mg/1 l-culture) by metal chelate chromatography. The fusion protein was cleaved with factor Xa, and the resulting rOAA was isolated in a final yield of 14.8 mg/1 l-culture by reversed-phase HPLC. Both the N-terminal sequence and the molecular mass of rOAA were found to be identical with those of OAA. The rOAA was fully functional with the same properties as OAA, as evidenced by hemagglutination activity, hapten-inhibition test, and binding specificity for high-mannose-type N-glycans. This rOAA should be applicable as a specific probe for high-mannose N-glycans and should contribute to elucidation of the molecular basis of its strict carbohydrate-binding specificity and potent anti-HIV activity.

Keywords

Anti-HIV High-mannose N-glycans Oscillatoria agardhii lectin Recombinant lectin 

Notes

Acknowledgments

We thank Dr Makoto Hirayama for his critical reading of the manuscript. We are grateful to Dr Haruyuki Iefuji for analyzing the N-terminal amino acid sequence of rOAA. This work was supported in part by a Grant-in-Aid for Scientific Research (B) from the Japan Society for the Promotion of Science.

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Copyright information

© The Japanese Society of Fisheries Science 2009

Authors and Affiliations

  1. 1.Graduate School of Biosphere ScienceHiroshima UniversityHiroshimaJapan

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