Purification and multimer formation of allurin, a sperm chemoattractant from Xenopus laevis egg jelly

Hitoshi Sugiyama, Lindsey Burnett, Xueyu Xiang, John Olson, Shaun Willis, Amy Miao, Tatsuo Akema, Allan L. Bieber, Douglas E. Chandler

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Allurin, a sperm chemoattractant isolated from Xenopus laevis egg jelly, can be purified in one step from an extract of diffusible jelly proteins ("egg water") using a FPLC or HPLC anion exchange column and a multi-step NaCl gradient. Allurin homomultimers were detected by Western blotting with antibodies prepared against the purified protein or peptides within the protein. Allurin multimers were stable and resisted dissociation by SDS and β-mercaptoethanol. Alkylation of allurin provided evidence for two free sulfhydryl groups but did not eliminate multimer formation, suggesting that intermolecular disulfide bond formation is not required for allurin aggregation. Concentration of egg water was accompanied by a reduction of chemoattractant activity that could not be fully accounted for by homomultimer formation. Rather, the presence of a multiphasic dose-activity curve upon partial purification and formation of hetero-allurin complexes during concentration suggested that egg water may contain allurin-binding proteins that reduce multimer formation and activity.

Original languageEnglish (US)
Pages (from-to)527-536
Number of pages10
JournalMolecular reproduction and development
Volume76
Issue number6
DOIs
StatePublished - Jun 1 2009

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology
  • Cell Biology

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