Designing of peptides with immuno-modulatory properties using protein A as a probe.

A series of reports from our laboratory have described the multifarious properties of protein A of Staphylococcus aureus Cowan I, apart from its IgG binding affinity. Original reports regarding its anti-tumor, anti-toxic, anti-carcinogenic and immunomodulatory properties published earlier by the authors have implicated some uniqueness of this bacterial protein. It was conceived that such diversified properties must lie in its specific peptide sequences, rendering it to act and behave as a multipotent "Biological Response Modifier" (BRM). The high resolution X-ray structure of protein A-Fc complex has been delineated earlier, and has been the foundation of many protein engineering studies. This structure along with the amino acid sequence data of its four repetitive domains provided us the basis for designing an octapeptide. This octapeptide was synthesized by solid phase peptide synthesis considering it as the probable site through which PA binds IgG. This octapeptide (NH2-Gln-Asn-Ala-Phe-Tyr-Glu-Ile-Leu-COOH) is present in the first helical segment of B-domain of protein A, and also is a part of domain D, A and C. This octapeptide has been shown to bind IgG by the immunoblotting technique. The binding affinity of the octapeptide appears to be significantly higher than that of intact protein A, as was revealed by calculation of Ka (association constant) and Kd (dissociation constant) values. This octapeptide might serve as a good immunoadsorbant for IgG and/or immune complexes.

In vitro anti-oxidant property of protein-A of Staphylococcus aureus.

Protein-A (PA) is a cell-surface glycoprotein of S. aureus Cowan I with immunomodulatory and anti-tumor activities, and ability to ameliorate cyclophosphamide and carbontetrachloride (CCl4) induced toxicity in rodents. The likely mechanism of this effect appears to be the anti-oxidant property of PA, evidenced in the present study by inhibition of CCl4 and Fe2-ascorbate induced lipid peroxidation in rat liver homogenates and inhibition of deaminative-oxidative degradation of L-glutamate into 2-thiobarbituric acid reactive products in a constituted chemical system. The anti-oxidant property of PA seem to arise from its molecular characteristics and the ability to interact with a superoxide derived free-radical species without any affinity for superoxide anion, hydroxyl radical and singlet oxygen species.