Albumin is synthesized in epididymis and aggregates in a high molecular mass glycoprotein complex involved in sperm-egg fertilization

Autor: Stephen Hyslop, Hiromitsu Tanaka, Kelen Fabiola Arroteia, Mainara Ferreira Barbieri, Luis Antonio Violin Dias Pereira, Marcos N. Eberlin, Lúcia Elvira Alvares, Gustavo H.M.F. Souza
Jazyk: angličtina
Rok vydání: 2014
Předmět:
Male
Physiology
lcsh:Medicine
Biochemistry
Epitope
Analytical Chemistry
Mice
Sperm-Egg Interactions
Reproductive Physiology
Medicine and Health Sciences
Zona pellucida
lcsh:Science
reproductive and urinary physiology
Epididymis
chemistry.chemical_classification
Multidisciplinary
Chromatographic Techniques
Antibodies
Monoclonal
Spermatozoa
Chemistry
medicine.anatomical_structure
Physical Sciences
Electrophoresis
Polyacrylamide Gel
Female
Anatomy
Research Article
Spectrometry
Mass
Electrospray Ionization
Urology
Immunoblotting
Biology
Research and Analysis Methods
Glycoprotein complex
Albumins
Column Chromatography
medicine
Animals
Immunoprecipitation
Glycoproteins
Ovum
Plasma Proteins
Molecular mass
lcsh:R
Reproductive System
Albumin
Biology and Life Sciences
Proteins
Molecular biology
Sperm
Mice
Inbred C57BL
chemistry
Fertilization
Infertility
lcsh:Q
Glycoprotein
Developmental Biology
Zdroj: PLoS ONE, Vol 9, Iss 8, p e103566 (2014)
PLoS ONE
ISSN: 1932-6203
Popis: The epididymis has an important role in the maturation of sperm for fertilization, but little is known about the epididymal molecules involved in sperm modifications during this process. We have previously described the expression pattern for an antigen in epididymal epithelial cells that reacts with the monoclonal antibody (mAb) TRA 54. Immunohistochemical and immunoblotting analyses suggest that the epitope of the epididymal antigen probably involves a sugar moiety that is released into the epididymal lumen in an androgen-dependent manner and subsequently binds to luminal sperm. Using column chromatography, SDS-PAGE with in situ digestion and mass spectrometry, we have identified the protein recognized by mAb TRA 54 in mouse epididymal epithelial cells. The ∼65 kDa protein is part of a high molecular mass complex (∼260 kDa) that is also present in the sperm acrosomal vesicle and is completely released after the acrosomal reaction. The amino acid sequence of the protein corresponded to that of albumin. Immunoprecipitates with anti-albumin antibody contained the antigen recognized by mAb TRA 54, indicating that the epididymal molecule recognized by mAb TRA 54 is albumin. RT-PCR detected albumin mRNA in the epididymis and fertilization assays in vitro showed that the glycoprotein complex containing albumin was involved in the ability of sperm to recognize and penetrate the egg zona pellucida. Together, these results indicate that epididymal-derived albumin participates in the formation of a high molecular mass glycoprotein complex that has an important role in egg fertilization.
Databáze: OpenAIRE
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