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Bovine seminal plasma osteopontin: Structural modelling, recombinant expression and its relationship with semen quality
Author(s) -
Cunha BustamanteFilho Ivan,
Renato Menegassi Silvio,
Ribas Pereira Gabriel,
Dias Salton Gabrielle,
Mosena Munari Fernanda,
Roberto Schneider Marlon,
Costa Mattos Rodrigo,
Otávio Jardim Barcellos Júlio,
Pereira Laurino Jomar,
Obino CirneLima Elizabeth,
Inês Mascarenhas Jobim Maria
Publication year - 2021
Publication title -
andrologia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.633
H-Index - 59
eISSN - 1439-0272
pISSN - 0303-4569
DOI - 10.1111/and.13905
Subject(s) - osteopontin , cd44 , recombinant dna , glycosaminoglycan , biology , sperm , integrin , hyaluronic acid , semen , capacitation , andrology , chemistry , microbiology and biotechnology , biochemistry , immunology , cell , gene , genetics , medicine
Osteopontin (OPN) is a multifunctional phosphoprotein that has been linked to fertility in bulls. However, the exact mechanism by which OPN contributes to fertilisation is yet unknown. The biotechnological use of OPN in bovine reproduction is promising but some gaps remain unfilled. The present work aimed: (a) to verify whether the seminal plasma OPN is associated with seminal traits and a standard breeding soundness exam; (b) to predict OPN interactions with integrins, CD44 and glycosaminoglycans through molecular docking; and (c) to develop a protocol for recombinant expression of OPN from vesicular gland cDNA. Ejaculates from top ranked bulls had higher amounts of seminal plasma OPN in comparison with bulls classified as questionable ( p < .01). The structural modelling and molecular docking predictions indicated that bovine OPN binds to heparin disaccharide, hyaluronic acid and hyaluronan. In addition, docking studies described the binding complexes of OPN with CD44 and the integrin heterodimers α5β1 and αVβ3. Finally, expression of rOPN‐6His was successfully obtained after 3 hr of induction with 0.5 mM IPTG at 37°C and a denaturing purification protocol resulted in efficiently purified recombinant OPN. The present results contribute to the development of biotechnological uses of OPN as a biomarker in bovine reproduction.