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Molecular cloning and expression of a rat homologue of human monocyte chemoattractant protein-3 (MCP-3) were studied. The rat MCP-3 gene was found in a 439-bp fragment by reverse transcription-polymerase chain reaction (RT-PCR) using degenerate primers based on conserved regions among other chemokines. Rat MCP-3 cDNA is 925 bp in length and contains an open reading frame of 284 amino acids. Its amino acid sequence had a high homology to the human and mouse MCP-3 sequences, although a cysteine residue is present at the position corresponding to the second cysteine residue in the human and mouse sequences. When MCP-3 was expressed in CHO cells, it was secreted to the cell culture supernatant, which was further purified by affinity chromatography. Rat MCP-3 also activated guinea pig neutrophils and human monocytes. The expression of the MCP-3 receptor was examined in a rat brain. Antibodies against the rat MCP-3 receptor recognized a 40 kDa protein in the membrane fraction of brain, suggesting that the MCP-3 receptor may exist in a proteolytically modified form on the cell surface.Q:
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