Recombinant expression of porcine spermadhesin AWN and its phospholipid interaction: Indication for a novel lipid binding property.
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.
Department Reproduction Biology, Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany.
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Schröter
Department Reproduction Biology, Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany.
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K
Müller
Department of Biology/Biophysics, Humboldt University of Berlin, Berlin, Germany.
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P
Müller
Department Mass Spectrometry, Leibniz Institute for Molecular Pharmacology, Berlin, Germany.
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E
Krause
Department Reproduction Biology, Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany.
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Braun
D000818:Animals; D002308:Cardiolipins / Q000378:metabolism; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000302:isolation & purification*; D002850:Chromatography, Gel / Q000662:veterinary; D004926:Escherichia coli / Q000378:metabolism; D008297:Male; D010712:Phosphatidic Acids / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism*; D012661:Semen; D029589:Seminal Plasma Proteins / Q000096:biosynthesis* / Q000302:isolation & purification*; D013552:Swine
D002308:Cardiolipins; D002352:Carrier Proteins; D010712:Phosphatidic Acids; D011994:Recombinant Proteins; D029589:Seminal Plasma Proteins; C425042:spermadhesin