Identification of the sperm-activating factor initiatorin, a prostatic endopeptidase of the silkworm, Bombyx mori

Insect Biochemistry and Molecular Biology
Sumiharu NagaokaKaeko Kamei

Abstract

Male Bombyx mori has a trypsin-type protease, called initiatorin, in the secretion from the posterior segment of the ejaculatory duct that is thought to be involved in the acquisition of sperm motility, although this inference remains to be demonstrated. Here, we revised the experimental procedures including that for purification and definitely identified the purified initiatorin protein as an activation factor of B. mori sperm by an in vitro study in which we treated isolated spermatozoa with this enzyme. Analysis of cDNA revealed that initiatorin consists of 281 amino acids with sequence similarity to bovine trypsin, and is highly homologous to the ejaculated accessory gland proteins not only of other Lepidoptera but also of Orthoptera. Recombinant initiatorin, expressed in Escherichia coli and purified, also showed proteolytic and sperm-activating activities. RT-PCR and Western blot analyses indicated that initiatorin is abundantly expressed in the glandula (g.) prostatica. It was also shown that pro-initiatorin is synthesized and stored in g. prostatica, and then converted to the mature form upon ejaculation. Fluorogenic peptides with a dibasic sequence were efficiently cleaved by initiatorin, and one such substrate, BOC-Gl...Continue Reading

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Citations

Oct 31, 2012·Molecular Reproduction and Development·Brooke A Laflamme, Mariana F Wolfner
Mar 27, 2018·Molecular Reproduction and Development·Kimberly StephensCatherine D Thaler
Dec 16, 2018·Molecular & Cellular Proteomics : MCP·Ethan C DegnerSteve Dorus
Mar 15, 2019·Molecular & Cellular Proteomics : MCP·Ethan C DegnerSteve Dorus
Feb 3, 2021·Insects·Rong-Er ZhengYipeng Xu
Feb 15, 2021·Journal of Insect Physiology·Julian G Shepherd, Karen Sartoris Bonk

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