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Recombinant Expression of Goat Milk Serum Amyloid A: Preliminary Studies of the Protein and Derived Peptides on Macrophage Phagocytosis

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Serum Amyloid A3 (SAA3) protein is a member of a complex group of acute phase and constitutive proteins which have been related to several immune functions. Bovine milk SAA3 (M-SAA3) has been described to have a unique N-terminal TFLK motif responsible for up regulating mucin expression in the intestine lumen and therefore a protective gastrointestinal role. cDNA sequences encoding the protein goat M-SAA3 were successfully cloned from milk, mammary gland tissue and liver, expressed despite observed toxicity and purified as a soluble protein. Sequence analyses of the milk and liver derived forms revealed a non mammary-restricted common N-terminal TFLR motif, unlike that described for bovine M-SAA3. Serum derived forms of SAA have been described to opsonize Gram-negative bacteria facilitating their phagocytosis by circulating macrophages or intestinal epithelial cells. However, no reports about a possible opsonic mechanism of the SAA3 isoforms have been described. Recombinant protein but not peptides encompassing the TFLR region increased blood and milk macrophage interaction and uptake of bacteria reported as number of bacteria per 100 macrophages and percentage of macrophages containing one or more bacteria. gMSAA3-derived peptides did not show any effect on phagocytosis. This would indicate that the TFLK-like region responsible for the up-regulation of mucins in the intestine is not the functional part of g-MSAA3 in promoting macrophage phagocytosis.

Keywords: Goat; Gram-negative bacteria; SAA3; epithelial cells; gMSAA3-derived peptides; macrophage; milk; opsonize; phagocytosis; serum amyloid A

Document Type: Research Article


Publication date: March 1, 2012

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  • Protein & Peptide Letters publishes short papers in all important aspects of protein and peptide research, including structural studies, recombinant expression, function, synthesis, enzymology, immunology, molecular modeling, drug design etc. Manuscripts must have a significant element of novelty, timeliness and urgency that merit rapid publication. Reports of crystallisation, and preliminary structure determinations of biologically important proteins are acceptable. Purely theoretical papers are also acceptable provided they provide new insight into the principles of protein/peptide structure and function.

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