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Reproductive Biology and Endocrinology
Published in: Reproductive Biology and Endocrinology 1/2015

Open Access 01-12-2015 | Research

Functional characterization of the domains of the bovine binder of SPerm 5 (BSP5) protein

Authors: Prashanth Sirigeri Jois, Geneviève Plante, Isabelle Thérien, Puttaswamy Manjunath

Published in: Reproductive Biology and Endocrinology | Issue 1/2015

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Abstract

Background

Bovine BSP5 is a multifunctional protein primarily involved in sperm capacitation. BSP5 consists of long N-terminal part followed by two similar and highly conserved fibronectin type II domains designated A and B.

Methods

In order to assess the role of these domains in the sperm binding and capacitation processes, we created recombinant individual domains (N, A, B), series of overlapping domains (NA and AB) and full-length BSP5 in an Escherichia coli expression system. The recombinant constructs were also tested for their ability to interact with ligands such as gelatine, heparin, chondroitin sulphate B and phosphatidylcholine liposomes by affinity chromatography and co-sedimentation studies.

Results

With the exception of the N domain, all recombinant constructs retained gelatine, phosphatidylcholine, heparin and chondroitin sulphate B binding activities. Domain-wise studies showed clearly that AB domain is capable of performing its biological functions as well as the full-length protein, as it was able to potentiate heparin-mediated sperm capacitation.

Conclusions

These results indicate that the C-terminal domain composed of two Fn2 domains is sufficient and crucial to maintain the biological functions of BSP proteins. The N-terminal part of the protein did not bind to any of known BSP5-ligands including epididymal sperm and did not seem to be required for either sperm binding or sperm capacitation. This study also confirmed that glycosylation is not required for BSP-mediated sperm capacitation or any of the binding characteristics displayed by BSP5.
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Metadata
Title
Functional characterization of the domains of the bovine binder of SPerm 5 (BSP5) protein
Authors
Prashanth Sirigeri Jois
Geneviève Plante
Isabelle Thérien
Puttaswamy Manjunath
Publication date
01-12-2015
Publisher
BioMed Central
Published in
Reproductive Biology and Endocrinology / Issue 1/2015
Electronic ISSN: 1477-7827
DOI
https://doi.org/10.1186/s12958-015-0058-4

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