Recombinant expression of porcine spermadhesin AWN and its phospholipid interaction: Indication for a novel lipid binding property

Abstract

AWN is a porcine (Sus scrofa domestica) seminal plasma protein and has been linked to a variety of processes related to fertilization. To acquire the protein in sufficient amount and purity for functional studies, we established its recombinant expression in E. coli and a three-step purification protocol based on different chromatographies. The test for AWN-phospholipid interaction revealed phosphatidic acid and cardiolipin as potential binding partners. As phosphatidic acid is surmised to play a role in cation-induced membrane destabilization and fusion events, we propose a membrane protective function of the presented binding affinity. Further studies with recombinant AWN will allow new insights into the mechanism of sperm-spermadhesin interaction and might provide new approaches for artificial reproduction techniques.

Protein signatures of seminal plasma from bulls with contrasting frozen-thawed sperm viability

“…Spermadhesins also share amino acid homology with seminal plasma motility inhibitor, a molecule that interferes with ATPase and reduces sperm motility in humans and boars 83 . However, studies demonstrate that phospholipid binding attributes of AWN, a porcine spermadhesin, promote stability of the sperm membrane 84 . It seems possible, then, that spermadhesins can have either beneficial or damaging effects on sperm, depending on the concentration.…”

“…Sperm capacitation; sperm interaction with the oviduct epithelium; sperm-egg binding; sperm membrane stability and motility. Anti-oxidative effects46,47,48,81,82,83,84…”

Bovine seminal plasma osteopontin: Structural modelling, recombinant expression and its relationship with semen quality

“…Despite the use of different vector and expression system, our results are similar as those found by (Bissonnette et al, 2012), who expressed recombinant OPN from cDNA obtained in from mammary gland tissue. So far, few seminal proteins from different domestic species were successfully expressed using heterologous systems including spermadhesins from bovine (aSFP (Bustamante-Filho et al, 2014), caprine (bodhesin, (Nascimento et al, 2012)) and swine (AWN (Schroter et al, 2017)) and binding of sperm proteins (BSPs) from bovine (PDC-109 (Singh et al, 2019) and human (BSPH1 (Lefebvre et al, 2009)). The production of recombinant seminal plasma proteins is a promising tool to create new reproductive biotechnologies, such seminal plasma proteins quantification methods and the development of semen extenders supplemented with recombinant protein (Zalazar et al, 2016).…”