Oocyte specific oolemmal SAS1B involved in sperm binding through intra-acrosomal SLLP1 during fertilization

Monika Sachdev, Arabinda Mandal, Sabine Mulders, Laura C. Digilio, Subbarayalu Panneerdoss, Viswanadhapalli Suryavathi, Eusebio Pires, Kenneth L. Klotz, Laura Hermens, María Belén Herrero, Charles J. Flickinger, Marcel van Duin, John C. Herr

Research output: Contribution to journalArticle

32 Scopus citations

Abstract

Molecular mechanisms by which fertilization competent acrosome-reacted sperm bind to the oolemma remain uncharacterized. To identify oolemmal binding partner(s) for sperm acrosomal ligands, affinity panning was performed with mouse oocyte lysates using sperm acrosomal protein, SLLP1 as a target. An oocyte specific membrane metalloproteinase, SAS1B (Sperm Acrosomal SLLP1 Binding), was identified as a SLLP1 binding partner. cDNA cloning revealed six SAS1B splice variants, each containing a zinc binding active site and a putative transmembrane domain, with signal peptides in three variants. SAS1B transcripts were ovary specific. SAS1B protein was first detected in early secondary follicles in day 3 ovaries. Immunofluorescence localized SAS1B to the microvillar oolemma of M2 oocytes. After fertilization, SAS1B decreased on the oolemma and became virtually undetectable in blastocysts. In transfected CHO-K1 cells SAS1B localized to the surface of unpermeabilized cells. Recombinant and native SLLP1 co-localized with SAS1B to the microvillar domain of ovulated M2 oocytes. Molecular interactions between mouse SLLP1 and SAS1B were demonstrated by surface plasmon resonance, far-western, yeast two-hybrid, recombinant- and native-co-IP analyses. SAS1B bound to SLLP1 with high affinity. SAS1B had protease activity, and SAS1B protein or antibody significantly inhibited fertilization. SAS1B knockout female mice showed a 34% reduction in fertility. The study identified SAS1B-SLLP1 as a pair of novel sperm-egg binding partners involving the oolemma and intra-acrosomal compartment during fertilization.

Original languageEnglish (US)
Pages (from-to)40-51
Number of pages12
JournalDevelopmental Biology
Volume363
Issue number1
DOIs
StatePublished - Mar 1 2012
Externally publishedYes

Keywords

  • Fertilization
  • Oocyte
  • SAS1B
  • SLLP1
  • Sperm
  • Sperm-egg binding

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

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  • Cite this

    Sachdev, M., Mandal, A., Mulders, S., Digilio, L. C., Panneerdoss, S., Suryavathi, V., Pires, E., Klotz, K. L., Hermens, L., Herrero, M. B., Flickinger, C. J., van Duin, M., & Herr, J. C. (2012). Oocyte specific oolemmal SAS1B involved in sperm binding through intra-acrosomal SLLP1 during fertilization. Developmental Biology, 363(1), 40-51. https://doi.org/10.1016/j.ydbio.2011.12.021