Transglutaminase activity in normal and hereditary cataractous rat lens and its partial purification.
Hereditary cataractous rat lenses showed significantly higher specific activity for transglutaminase than the normal lenses of comparable age. Transglutaminase activity of normal lenses was distributed predominantly in the buffer-soluble fraction. The buffer-insoluble fraction showed 14% of total activity. The cortical and nuclear fractions of the normal lens showed 43% and 57% distribution of total activity, respectively. Protein solubilizing agents enhanced the activity of the enzyme in the lens homogenate in the following order: Triton X-100 greater than Tween 20 greater than Sodium dodecyl sulfate greater than Sodium deoxycholate greater than Sodium cholate greater than NaSCN greater than KI. Transglutaminase was purified 15 fold by hydrophobic affinity chromatography employing omega-amine octylagarose matrix. The purified enzyme was activated by calcium and inactivated by iodoacetamide and upon freeze-drying. Lens crystallins served as exogenous substrate for transglutaminase, with gamma-crystallin as the most effective amine acceptor.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002386:Cataract / Q000201:enzymology*; D002793:Cholic Acids / Q000494:pharmacology; D003840:Deoxycholic Acid / Q000494:pharmacology; D007460:Iodoacetamide / Q000494:pharmacology; D007908:Lens, Crystalline / Q000187:drug effects / Q000201:enzymology*; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology; D011136:Polysorbates / Q000494:pharmacology; D011193:Potassium Iodide / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013861:Thiocyanates / Q000494:pharmacology; D005723:gamma-Glutamyltransferase / Q000032:analysis*
D002793:Cholic Acids; D011136:Polysorbates; D013861:Thiocyanates; D003840:Deoxycholic Acid; D011193:Potassium Iodide; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D017830:Octoxynol; D005723:gamma-Glutamyltransferase; D002118:Calcium; D007460:Iodoacetamide