Functional mimicry of protein A of Staphylococcus aureus by a proteolytically cleaved fragment.
Protein A (PA) of Staphylococcus aureus has an array of biological functions, such as antitumor, antitoxic, anticarcinogenic, immunomodulatory, antifungal, and antiparasitic properties. We have already established that a theoretical trypsin-digested peptide fragment of protein A (20-mer) mimics immunomodulatory and IgG binding property of PA. In the present report we have concentrated on a 16-mer chymotryptic fragment of protein A, which has a sequence of 13 amino acids in common with the previously studied 20-mer peptide. Molecular modeling study qualitatively predicted that both 20-mer and 16-mer peptides retain Fc binding ability from an interaction energy point of view. In the present study our aim was to understand whether this theoretically predicted 16-mer chymotryptic fragment could be formed in a real experiment and also to understand its biological activities. Chymotrypsin cleavage of PA at 37 degrees C for 24 h produced four major fragments on reverse-phase HPLC. The amino acid analyses of each fragment show the absence of cysteine residue from all fragments, which justifies the absence of cysteine in PA. We also observed high content of aspartic acid and glutamic acid residues in all fragments. On gel-filtration chromatography the chymotrypsin cleavage of PA shows five peaks, one of which overlaps with our theoretically selected 16-mer peptide on superimposition. We verified the IgG binding capacity of 16-mer peptide by capillary electrophoresis. The 16-mer peptide also induces the production of TNFalpha and IL-1alpha in serum of mice. The above observations suggest that the 16-mer peptide may be produced by chymotrypsin cleavage and also that this peptide possesses some of the major biological properties of PA, such as IgG binding, TNFalpha and IL-1alpha elicitation, etc.
Sinha
P
P
Immunotechnology Section, Bose Institute, Calcutta, 700 054, India.
Sengupta
J
J
Ray
P K
PK
D000595:Amino Acid Sequence; D000818:Animals; D002850:Chromatography, Gel; D002918:Chymotrypsin / Q000494:pharmacology; D006801:Humans; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D017452:Receptors, IgG / Q000737:chemistry; D013205:Staphylococcal Protein A / Q000737:chemistry* / Q000276:immunology; D013997:Time Factors
D010455:Peptides; D017452:Receptors, IgG; D013205:Staphylococcal Protein A; D002918:Chymotrypsin