Isolation and characterization of a 25-kilodalton protein from mouse testis: sequence homology with a phospholipid-binding protein.
A 25-kDa epididymal secretory protein (MEP 9), isolated from mouse epididymal fluid, has recently been characterized in our laboratory [Rankin et al., Biol Reprod 1992; 46:747-766]. The polyclonal antibody raised against this protein was found to recognize a 25-kDa component in epididymal fluid and testicular extract. The 25-kDa testicular antigen (MTP) was purified by means of ammonium sulfate precipitation, gel filtration, and anion-exchange chromatography; MTP was found to be similar to MEP 9 in several properties including molecular mass (25 kDa), isoelectric point (pI 6.0), and immunoreactivity when the proteins were resolved in the presence of SDS (one-dimensional and two-dimensional PAGE). However, when the proteins were resolved under non-denaturing conditions, MTP showed strong immunoreactivity while MEP 9 did not. This observation suggests that although the 25-kDa antigens from the epididymal fluid and testicular extract are quite similar, they may have different immunological conformations. When analyzed for amino acid composition and partial amino acid sequence, the testicular antigen showed substantial homology (> 80%) with a phosphatidylethanolamine-binding protein characterized from bovine brain. MTP also showed phosphatidylethanolamine-binding activity (Kd = 1.95 x 10(-5) M, Bmax = 1.86 nmol/micrograms MTP), suggesting that the mouse 25-kDa protein is a member of the phospholipid-binding protein family and may have a role in lipid metabolism during sperm maturation.
Araki
Y
Y
Center for Reproductive Biology Research, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-2633.
Vierula
M E
ME
Rankin
T L
TL
Tulsiani
D R
DR
Orgebin-Crist
M C
MC
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D050356:Lipid Metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D010714:Phosphatidylethanolamines / Q000378:metabolism; D011506:Proteins / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification*; D017386:Sequence Homology, Amino Acid; D013737:Testis / Q000378:metabolism*
C079124:MEP 9 protein, mouse; D010714:Phosphatidylethanolamines; D011506:Proteins; C079125:testicular antigen, 25 kDa