Proteomics of Tribolium castaneum seminal fluid proteins: Identification of an angiotensin-converting enzyme as a key player in regulation of reproduction

Seminal fluid proteins (SFPs) play important roles in regulation of reproduction and behavior. Proteomics approaches were used to identify 13 SFPs, including 3 new proteins in the red flour beetle, Tribolium castaneum. The 13 SFP genes identified code for Serpin, cysteine-rich protein, odorant binding protein-like (OBPL, G10064 and G10065), Kunitz-like protease inhibitor precursor, and WD 40 family protein and are predominantly expressed in the male accessory glands. The genes coding for 13 putative SFPs were knocked down in males; the RNAi males were mated with virgin females, and the number of eggs produced by the mated females was quantified. Knockdown in the expression of the gene coding for a protein similar to angiotensin-converting enzyme 9 (G15465, TcACE) in the males caused a decrease in egg production by the females when compared to the eggs produced by the females mated with control males. In addition, knockdown in the expression of the gene coding for heat shock cognate 70 led to a reduction in the amount of proteins produced by the male accessory glands by 55%. These data suggest that angiotensin-converting enzyme produced in the male seminal vesicles plays important roles in sperm protection during and after transfer to females.