title: PROPERTIES OF PROTEINASE FROM STREPTOCOCCUS FAECALIS VAR. LIQUEFACIENS.
abstract: Bleiweis, Arnold S. (The Pennsylvania State University, University Park), and Leonard N. Zimmerman. Properties of proteinase from Streptococcus faecalis var. liquefaciens. J. Bacteriol. 88:653-659. 1964.-The extracellular group D streptococcal proteinase is inactivated by chelating agents [ethylenediamine-tetraacetate (EDTA), o-phenanthroline, and 8-quinolinol] and mercaptans (cysteine, mercaptoethanol, and thioglycolate). The optimal inhibitory concentrations of EDTA (4 x 10(-4)m) and cysteine (2.5 x 10(-2)m) promote rapid loss of activity with 50% inactivation after 4 to 5 min. Enzyme inactivated by either EDTA or cysteine is reactivated about 80% by 4 x 10(-4)m Zn(++). Such reactivation of EDTA-treated enzyme is prevented completely by iodoacetate (5 x 10(-2)m) and of cysteine-treated enzyme by oxidizing conditions, which suggests that the zinc binding-site may be a thiol. High levels of zinc (10(-3)m) do not allow reactivation in either case, and actually inhibit native proteinase. Ca(++), Mg(++), Co(++), Fe(++), Cu(++), and Ni(++) do not reactivate cysteine-treated enzyme, but Mn(++) (10(-4) to 8 x 10(-4)m) allows 27% reversal. N(2)-held, cysteine-treated enzyme can be spontaneously reactivated if the substrate is flushed with O(2) during the assay; leakage of air or O(2) into the samples before assay leads to loss of reactivatability. Native proteinase does not lose activity after dialysis for 43 hr against 0.07 m phosphate buffer. It is concluded that the group D proteinase obtained from Streptococcus faecalis var. liquefaciens is probably a zinc metalloenzyme that is unlike the thiol-activated, group A streptococcal proteinase, but similar to the mammalian carboxypeptidase A.
mesh_terms: D002118:Calcium; D002614:Chelating Agents; D003035:Cobalt; D003300:Copper; D003545:Cysteine; D004492:Edetic Acid; D010450:Endopeptidases; D013293:Enterococcus faecalis; D007461:Iodoacetates; D007501:Iron; D008274:Magnesium; D008623:Mercaptoethanol; D009532:Nickel; D010447:Peptide Hydrolases; D010600:Pharmacology; D010618:Phenanthrolines; D011804:Quinolines; D006435:Renal Dialysis; D013864:Thioglycolates; D015032:Zinc
chemical_list: D002614:Chelating Agents; D007461:Iodoacetates; D010618:Phenanthrolines; D011804:Quinolines; D013864:Thioglycolates; D003035:Cobalt; D008623:Mercaptoethanol; D003300:Copper; D009532:Nickel; D004492:Edetic Acid; D007501:Iron; D010450:Endopeptidases; D010447:Peptide Hydrolases; D008274:Magnesium; D015032:Zinc; D003545:Cysteine; D002118:Calcium; C025205:1,10-phenanthroline
keywords: CALCIUM; COBALT; COPPER; CYSTEINE; IODOACETATES; IRON; MAGNESIUM; MERCAPTOETHANOL; NICKEL; PEPTIDE HYDROLASES; PHARMACOLOGY; PHENANTHROLINES; QUINOLINES; STREPTOCOCCUS FAECALIS; THIOGLYCOLATES; ZINC