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Galectin-3 is associated with prostasomes in human semen

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Abstract

Galectin-3 is a β-galactoside-binding protein involved in immunomodulation, cell interactions, cancer progression, and pathogenesis of infectious organisms. We report the identification and characterization of galectin-3 in human semen. In the male reproductive tract, the ~30 kDa galectin-3 protein was identified in testis, epididymis, vas deferens, prostate, seminal vesicle, and sperm protein extracts. In seminal plasma, galectin-3 was identified in the soluble fraction and in prostasomes, cholesterol-rich, membranous vesicles that are secreted by the prostate and incorporated into seminal plasma during ejaculation. Two-dimensional immunoblot analysis of purified prostasomes identified five galectin-3 isoelectric variants with a pI range of 7.0 to 9.2. Affinity purification and tandem mass spectrometry of β-galactoside-binding proteins from prostasomes confirmed the presence of galectin-3 in prostasomes and identified a truncated galectin-3 variant. The intact galectin-3 molecule contains a carbohydrate recognition domain and a non-lectin domain that interacts with protein and lipid moieties. The identification of a monovalent galectin-3 fragment with conserved carbohydrate-binding activity indicates the functional relevance of this truncation and suggests a regulatory mechanism for galectin-3 in prostasomes. Surface biotinylation studies suggested that galectin-3 and the truncated galectin-3 variant are localized to the prostasome surface. Prostasomes are proposed to function in immunosuppression and regulation of sperm function in the female reproductive tract, are implicated in facilitating sexually-transmitted infections, and are indicated in prostate cancer progression. Given the overlap in functional significance, the identification of galectin-3 in prostasomes lays the groundwork for future studies of galectin-3 and prostasomes in reproduction, disease transmission, and cancer progression.

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Abbreviations

BME:

β-mercaptoethanol

BSA:

bovine serum albumin

CRD:

carbohydrate recognition domain

Gal:

galactose

GalNAc:

N-acetylgalactosamine

GlcNAc:

N-acetylglucosamine

HRP:

horse radish peroxidase

kDa:

kilodaltons

PAGE:

polyacrylamide gel electrophoresis

PBS:

phosphate-buffered saline

pI:

isoelectric point

SDS:

sodium dodecyl sulfate

sulfo-NHS-LC-biotin:

sulfosuccinimidyl-6-(biotinamido) hexanoate

TBS:

Tris-buffered saline

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Acknowledgements

This work was funded by NIH HD50540 and a pilot study grant from the College of Medicine, University of Arkansas for Medical Sciences (UAMS) to A.B.D. The authors wish to thank the Proteomics Core Facility at UAMS.

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Author notes

  1. Cheryl F. Lichti

    Present address: Washington University School of Medicine, 660 S. Euclid Avenue, Campus Box 8127, St. Louis, MO, 63110, USA

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, College of Medicine, University of Arkansas for Medical Sciences, Little Rock, AR, 72205, USA

    Jennifer L. Jones, Sarika Saraswati, Ashley S. Block, Cheryl F. Lichti & Alan B. Diekman

  2. Department of Obstetrics and Gynecology, College of Medicine, University of Arkansas for Medical Sciences, Little Rock, AR, 72205, USA

    Maha Mahadevan

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  1. Jennifer L. Jones
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Correspondence to Alan B. Diekman.

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Jennifer L. Jones and Sarika Saraswati contributed equally to this article.

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Jones, J.L., Saraswati, S., Block, A.S. et al. Galectin-3 is associated with prostasomes in human semen. Glycoconj J 27, 227–236 (2010). https://doi.org/10.1007/s10719-009-9262-9

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  • DOI: https://doi.org/10.1007/s10719-009-9262-9

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