Chapter 16 the Citric Acid____ Cycle.pptVIP

Electron microscopy studies of a-keto acid dehydrogenase complexes Increasing resolution structures has been determined using EM. REED LJ, FERNANDEZ-MORAN H, KOIKE M, WILLMS CR. ELECTRON MICROSCOPIC AND BIOCHEMICAL STUDIES OF PYRUVATE DEHYDROGENASE COMPLEX OF ESCHERICHIA COLI. Science. 1964 Aug 28;145:930-2. Zhou ZH, McCarthy DB, OConnor CM, Reed LJ, Stoops JK. The remarkable structural and functional organization of the eukaryotic pyruvate dehydrogenase complexes. Proc Natl Acad Sci U S A. 2001 98:14802-7. E1 E2 E3 E1-Pyruvate dehydrogenase E2-Dihydrolipoyl transacetylase E3- Dihydrolipoyl dehydrogenase Elucidation of the glyoxylate cycle:background The intermediates of the citric acid cycle are obligatory precursors of many biosynthesis, thus will be continuously removed. Certain replenishment mechanism must exist. Isocitrate lyase was first discovered in 1953 (Campbell et al, BBA 11:594): catalyzing the formation of glyoxylate and succinate when citrate or cis-aconitate was added into bacterial extracts. Malate synthase was first revealed in 1956 (Wong and Ajl, JACS, 78:3230.): catalyzing the condensation of acetyl-CoA and glyoxylate to form malate. Elucidation of the glyoxylate cycle:Actual occurrence of a pathway Kornberg, H. L.; Madsen, N. B. (1957) Synthesis of C4-dicarboxylic acids from acetate by a glyoxylate bypass of the tricarboxylic acid cycle, Biochimica et Biophysica Acta, 24, 651-3. When cell-free exts. of acetate-grown Pseudomonas KB 1 were incubated with C14H3CO2Na (I), ATP, coenzyme A, glutathione, and Na glyoxylate, malate was the only labeled compound formed in the early stages of incubation. When isocitrate replaced glyoxylate, malate was again the 1st labeled compound formed. In the absence of glyoxylate or isocitrate, no labeled compds. were formed other than traces of acetyl CoA. It is concluded that Pseudomonas KB 1, growing on acetate as sole source of C, possesses, in addition to the enzymic reactions of the tricarboxylic acid