Gene expression and cDNA cloning identified a major basic protein constituent of bovine seminal plasma as bovine monocyte-chemoattractant protein-1 (MCP-1).
P6 is one of the major basic proteins of bovine seminal plasma. Using cell-free translation of poly(A)+RNA from bovine seminal vesicle tissue and monospecific anti-P6-IgGs, we show that P6 is a secretory product of the seminal vesicles. Immunohistochemical experiments supported this finding. Immunoscreening of a lambda gt11 cDNA library derived from seminal vesicle poly(A)+RNA furnished a number of positive cDNA clones, from which clone pH42 was characterized by sequencing. The partial amino acid sequence of a CNBr-fragment of P6 permitted identification of the reading frame of clone pH42 encoding the precursor protein of P6. The P6 precursor contains a signal peptide of 23 amino acids followed by the mature P6 sequence of 76 amino acid residues. The cDNA sequence of pH42 was 80% homologous with that of the human monocyte-chemoattractant protein-1 (hMCP-1). The respective amino acid sequences for the precursor molecules are 72% identical. Northern analysis of seminal vesicle poly(A)+RNA using pH42 as probe probe identified a 0.9-kb P6 mRNA. Stimulation of P6 mRNA expression by phytohemagglutinin in bovine peripheral mononuclear leukocytes suggests that P6 is identical to bovine MCP-1.
Wempe
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F
Max-Planck-Institut für Biophysikalische Chemie, Abt. Molekulare Biologie, Göttingen, Germany.
Henschen
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A
Scheit
K H
KH
D000595:Amino Acid Sequence; D000818:Animals; D000951:Antigens, Neoplasm / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002417:Cattle; D018932:Chemokine CCL2; D002630:Chemotactic Factors / Q000235:genetics*; D003001:Cloning, Molecular; D015870:Gene Expression; D015151:Immunoblotting; D008297:Male; D008969:Molecular Sequence Data; D011061:Poly A / Q000235:genetics; D029584:Prostatic Secretory Proteins; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics* / Q000302:isolation & purification; D012313:RNA / Q000235:genetics; D012333:RNA, Messenger; D029589:Seminal Plasma Proteins; D012669:Seminal Vesicles / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid
D000951:Antigens, Neoplasm; D018932:Chemokine CCL2; D002630:Chemotactic Factors; D029584:Prostatic Secretory Proteins; D011506:Proteins; D012333:RNA, Messenger; D029589:Seminal Plasma Proteins; C043076:beta-microseminoprotein; D011061:Poly A; D012313:RNA