Tyrosinases in Rana pipiens. Purification and physical properties.
Two enzymes with tyrosinase activity have been purified from the frog Rana pipiens. Both enzymes are isolated in an inactive form which can be activated with trypsin. Amino acid analysis, NH2-terminal amino acid determination (arginine for both proteins), and immunological evidence indicate that athe two enzymes are similar if not identical. They can be distinguished by their trypsin activation kinetics. Cell fractionation studies suggest that one form is found associated with the smooth endoplasmic reticulum whereas the other protein fraction is localized mainly within the premelanosomes. Sedimentation equilibrium studies demonstrate that both protein fractions are self-associating systmes. Ionic strength, temperature, and specific anion effects alter the equilibria of the associating systems. The monomeric molecule weight for both fractions is 30,000 and at low ionic strengths the predominant molecular weight species is the tetramer. The partial specific volume of each protein is 0.70.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D004156:Catechol Oxidase / Q000302:isolation & purification / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005779:Immunodiffusion; D007527:Isoenzymes / Q000302:isolation & purification; D008547:Melanophores / Q000201:enzymology; D008970:Molecular Weight; D011894:Rana pipiens; D012867:Skin / Q000201:enzymology*; D013347:Subcellular Fractions / Q000201:enzymology; D014357:Trypsin
D000596:Amino Acids; D007527:Isoenzymes; D004156:Catechol Oxidase; D014357:Trypsin