Streptococcal proteinase

An illustration of this approach may be seen in the studies on streptococcal proteinase (Liu 1967). The activity of this enzyme is dependent upon the presence of a free sulfhydryl group. The active form of the enzyme was first converted to the inactive S-sulfenyl-sulfonate derivative. Treatment of this derivative with a chemically-reactive substrate "analogue, a-N-bromoacetylarginine methyl ester, resulted in the alkylation of a single histidine residue. The sulfhydryl group of the modified enzyme was regenerated by reduction, however, this did not restore enzymatic activity, thus providing presumptive evidence for the involvement of both a cysteinyl and a histidyl residue in the active site of this enzyme. 

An alternative method for preparing derivatives of bromoacetate involves using bromoacetyl bromide or bromoacetyl chloride. The synthesis of the N-bromoacetyl derivative of L-arginine methyl ester with bromoacetyl bromide was carried out by Liu (1967). The product N-bromoacetyl-L-arginine methyl ester was a successful affinity label for streptococcal proteinase. L-Arginine methyl ester dihydrochloride (780 mg 3 mmoles) was dissolved in 6 ml of ice-cold 1 N NaHCOj. Over a period of 20 min, freshly distilled bromoacetylbromide, 3... 

Gerlach D, Knoll H, Kohler W, Ozegowski J, Hribalova V Isolation and characterization of erythrogenic toxins. 5. Communication identity of erythrogenic toxin type B and streptococcal proteinase precursor. Zentralbl Bakteriol Mikrobiol Hyg [A] 1983 255 221-233. 

Gerlach D, Reichardt W, Fleischer B, Schmidt K Separation of mitogenic and pyrogenic activities from so-called erythrogenic toxin type B (Streptococcal proteinase). Zentralbl Bakteriol 1994 280 507-514. 

Hauser, A.R. and Schlievert, P.M. 1990. Nucleotide sequence of the streptococcal pyrogenic exotoxin type B gene and toxin relationship to streptococcal proteinase precursor. J. Bacteriol. 172 4536-4542. 

Tai, J.Y., Kortt, A.A., Liu, T.Y., and Elliott, S.D. 1976. Primary structure of streptococcal proteinase. III. Isolation of cyanogen bromide peptides complete covalent structure of the polypeptide chain. J. Biol Chem. 251 1955-1959. 

Robinson, G. W. Reaction of a Specific Tryptophan Residue in Streptococcal Proteinase with 2-Hydroxy-5-nitrobenzyl Bromide. J. Biol. Chem. 245, 4832-4841... 

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