PURIFICATION AND CHARACTERIZATION OF A MALE-SPECIFIC PROTEIN IN THE HEMOLYMPH OF THE WAX MOTH, GALLERIA-MELLONELLA L

A male-specific protein (MSP) present only in males was identified fro m the hemolymph of the wax moth, Galleria mellonella L., by polyacryla mide gel electrophoresis (PAGE) and purified by anion-exchange chromat ography. MSP has a native molecular mass of 55 kDa and consists of two 27-kDa subunits. An isoelectric point of MSP was measured to be appro ximately 5.8. MSP is a glycoprotein that contains 1.7% carbohydrate. T he compositional analysis of carbohydrate component indicated a predom inance of fructose and glucose. MSP also contains large amounts of asp aragine, aspartic acid, glutamine, glutamic acid, and lysine but small amounts of tyrosine, methionine, and tryptophan. Western blot analysi s of the hemolymph of each developmental stage indicated that MSP is p resent in the hemolymph of 8 day-old pupa and adult. Also, results fro m Western blotting indicated that MSP is not present in the tissues of larvae and of female adults but appears in the fat body of male pupae and adult and testis of adult. The fat body and testis of male pupae and adult were cultured in vitro to trace the place and time of MSP sy nthesis. The fat body began to synthesize MSP in late pupae and showed active synthesis during the adult stage. The distribution of MSP in t he testis was observed by electron microscopic immunogold labeling, us ing the antibody against MSP. MSP is present between the germinal cyst s and is taken up through the basal surface of the seminiferous tubula r epithelium. (C) 1998 Wiley-Liss, Inc.