Analysis of transketolase and identification of an enzyme variant in human leukocytes.
Human leukocyte transketolase of fresh cell extracts has been analyzed by isoelectrofocusing on agarose gels (pH 3-10). The enzyme was then transblotted on nitrocellulose and detected with specific anti-transketolase IgG coupled to an avidin/biotin-immunoperoxidase system. Each sample yielded multiple enzyme forms, within a pI range of about 7.4-8.4. Transketolase profile, however, was not identical in all extracts. There are two mainly distinct patterns, showing qualitative and quantitative differences: a standard profile, which is predominant, and a variant, found in three unrelated subjects out of the two hundred and twenty. Standard and variant enzyme have similar Km values for ribose 5-P and xylulose 5-P and the same mobility on SDS-PAGE.
Paoletti
F
F
Istituto di Patologia Generale, Viale G.B. Morgagni, Firenze, Italy.
Mocali
A
A
Marchi
M
M
Truschi
F
F
D003600:Cytosol / Q000201:enzymology; D014644:Genetic Variation; D006801:Humans; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000097:blood / Q000235:genetics / Q000302:isolation & purification; D007962:Leukocytes / Q000201:enzymology*; D014174:Transketolase / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification
D007527:Isoenzymes; D014174:Transketolase