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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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