ISOLATION AND CHARACTERIZATION OF A 25-KILODALTON PROTEIN FROM MOUSE TESTIS - SEQUENCE HOMOLOGY WITH A PHOSPHOLIPID-BINDING PROTEIN

被引:32
作者
ARAKI, Y
VIERULA, ME
RANKIN, TL
TULSIANI, DRP
ORGEBINCRIST, MC
机构
[1] VANDERBILT UNIV, MED CTR,SCH MED,CTR REPROD BIOL RES, KIINAMYLLYNKATU 10, NASHVILLE, TN 37232 USA
[2] VANDERBILT UNIV, MED CTR, SCH MED, DEPT OBSTET & GYNECOL, NASHVILLE, TN 37232 USA
关键词
D O I
10.1095/biolreprod47.5.832
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
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 arc 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 (K(d) = 1.95 X 10(-5) M, B(max) = 1.86 nmol/mug 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.
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页码:832 / 843
页数:12
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