Separation of 2 Glutamic-Oxalacetic Transaminases by Paper Electrophoresis.

Abstract

of cwllagenous fillers, as seen for example in peridorit it is, results fro111 pririury alteratiori by the iriflariirnzatory dary attack by bacterial proteases.'I' r ea tm en t w it h ethylene oxicl c rendered reconstituted acid-soluble collagen readily digestible by colmibined pure cultures of human oral strains of fusiforrn bacilli, diphtheroid bacilli, streptococci, and veillonellae. The fact that only a few of these strains individually attacked such altered collagen to a slight extent indicates digestion by synergistic and symbiotic reactions between members of the oral microbiota. Concomitantly with increased digestibility from treatment with ethylene oxide, reconstituted collagen exhibited marked morphological changes as revealed by electron microscopy. S u jyc w? N v y . Mayo Clinic and M a y o Fn.,t Rochester, Ninn.The enzyme glutamic-oxalacetic transaminase (GOT) has attracted considerable interest recently because its concentrations in blood and tissue are altered characteristically in a numiber of disease states. Methods of purification devised up to now( 1,2) have produced no indication that this enzyme may occur in more than one form. We observed that crude GOT in an extract of cardiac muscle from dog, when chromatographed on certain ion exchangers, behaves as a mixture of 2 enzymes. When CM-cellulose was employed with phosphate buffer of pH 7.0 and r/2 = 0.01 or less, one component was adsorbed and could be eluted a t higher ionic strength and lower pH, while the other was adsorbed very little if at all. With DEAE-cellulose the be--*This investigation was supported in part by Research Grant from Nat. Tnst. of Arthritis and Metab. Diseases, P.H.S. t The Mayo Foundation, Rochester, Minn., is part of Graduate School of Univ. of Minnesota.havior olf the 2 fractions was reversed. These findings suggested that the 2 GOT'S may have opposite electrical charges at p H 7. To test this assumption, the paper electrophoresis of crude preparations of GOT was investigated.Methods and materials. Tissues were minced and extracted in a blendor with 3 volumes of cold water. The centrifuged extracts were concentrated several fold by ultrafiltration at 0°C. The solutions were recentrifuged at 30,000 g. They were then applied, in proportions of 10 to 40 pl, containing about 1 to 5 mg of protein, to the paper strips in a Spinco electrophoretic apparatus Model R. Phosphate buffer of pH 7.4, r/2 = 0.075, was used, often with the addition of one per cent bovine serum albumin as a stabilizer of enzymatic activity. Stabilization could be achieved also by applying quantities of tissue extracts corresponding to the larger amount of protein. Current was 5 milliamperes, temperature 8"C, at UNSW Library on July 18, 2015 ebm.sagepub.com Downloaded from

Lactate dehydrogenase and glutamic oxalic transaminase isozymes in heart muscle of forensic pathological cases.

“…Fleisher et al (1960) observed that GOTs/GOTm ratio of the heart muscle diminished as a consequence of myocardial infarction or acute suffocation. The death of myocardial cells increases the permeability of the cell membrane allowing GOT to pass out of the cell into the serum (Shikata et al 1970).…”

Clinical and Diagnostic Significance of Aspartate Aminotransferase Isoenzymes in Sera of Patients with Liver Diseases

“…Fleisher et al (3,4) found human serum to contain two types of aspartate aminotransferase. This finding was confirmed by subsequent enzymologic studies (2,5,6), which demonstrated the existence of a cytoplasmic or soluble isoenzyme of aspartate aminotransferase and a mitochondrial one.…”

“…22, 1984 / No. 2 partate aminotransferases (EC 2.6.1.1), (2)(3)(4)(5)(6), which differ in their amino acid composition and in their immunochemical and kinetics properties (7)(8)(9). As early äs 1967, Schmidt et al (10) started to consider the possible clinical usefulness of measuring of these isoenzymes.…”

Experimental model of sudden cardiac death from unexplained cause.

“…They are supernatant GOT (GOTs) and mitochondrial GOT (GOTm) (Fleisher et al 1960). In the heart muscle of the control group, GOTm and GOTs were about 57-73% and 31-41%, respectively, while GOTm of the experimental group decreased to about 49-66% of the total GOT.…”