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Biomedical subjects

Y F Chang

Publications and source records attributed to Y F Chang.

At least 127 records · Page 7Linked to original sources
PubMed1982

Lysine metabolism in the human and the monkey: demonstration of pipecolic acid formation in the brain and other organs.

Metabolism of L-[U-14C]lysine was studied in the human autopsy tissues and the intact monkeys through intracerebroventricular and intravenous injections. The human tissues were more active in the metabolism of L-[14C]lysine to [14C]pipecolate than the rat tissues previously reported. This metabolism was equally active in the phosphate (pH 7) and the glycyl-glycine (pH 8.6) buffers with the brain and the kidney having higher activity than the liver. Besides [14C]pipecolate, traces of [14C]saccharopine and alpha-[14C]aminoadipate were also detected in the liver incubation. Twenty-four hr after intraventricular injection of L-[14C]lysine to the monkey, substantial labeling of pipecolate and alpha-aminoadipate was observed in the brain and spinal cord, with the kidney, liver and the plasma having much reduced levels. Radioactivity levels of these two compounds were found low in the organs and plasma of the intravenously injected monkey. The urine of both monkeys contained only traces of [14C]pipecolate, even though it contained high levels of L-[14C]lysine and alpha-[14C]aminoadipate. It was concluded that L-lysine is actively metabolized to pipecolate and alpha-aminoadipate in the human and the monkey, that this reaction is most active in the brain when L-lysine is intraventricularly administered, and that in contrast to the rat, the monkey may have an effective renal reabsorption for pipecolate which is similar to the human.

2-Aminoadipic Acid↗
PubMed1982

delta1-piperideine-2-carboxylate reductase of Pseudomonas putida.

Pseudomonas putida metabolizes D-lysine to delta 1-piperideine-2-carboxylate and L-pipecolate. The second step of this catabolic pathway is catalyzed by delta 1-piperideine-2-carboxylate reductase. This enzyme was isolated and purified from cells grown on DL-lysine as substrate. The enzyme was very unstable, resulting in low recovery of activity and low purity after a six-step purification procedure. The enzyme had a pH optimum of 8.0 to 8.3. The Km values for delta 1-piperideine-2-carboxylate and NADPH were 0.23 and 0.13 mM, respectively. NADPH at concentrations above 0.15 mM was inhibitory to the enzyme. Delta 1-pyrroline-5-carboxylate, pyroglutamate, and NADH were poor substrates or coenzyme for delta 1-piperideine-2-carboxylate reductase. The enzyme reaction from delta 1-piperideine-2-carboxylate to L-pipecolate was irreversible. EDTA, sodium pyrophosphate, and dithiothreitol at concentrations of 1 mM protected the enzyme during storage. The enzyme was inhibited almost totally by Zn2+, Mn2+, Hg2+ Co2+, and p-chloromercuribenzoate at concentrations of 0.1 mM. The enzyme had a molecular weight of about 200,000. Both D-lysine and L-lysine were good inducers for the enzyme. Neither delta1-piperideine-2-carboxylate nor L-pipecolate was an effective inducer for the enzyme. P. putida cells grew on D-lysine only after a 5- to 8-h lag, which could be abolished by adding a supplement of 0.01% alpha-ketoglutarate or other readily metabolizable compounds. Such a supplement also converted the noncoordinate induction of this enzyme and pipecolate oxidase, both of the D-lysine pathway, to coordinacy. However, this effect was not observed if the enzyme pair was from different pathways of lysine metabolism in this organism (i.e., the D- and L-lysine pathways).

Culture Media↗
PubMed1981

Uptake, release, and metabolism of D- and L-alpha-aminoadipate by rat cerebral cortex.

Accumulation of L-alpha-aminoadipate by rat cerebral cortical slices is a stereospecific and Na+-dependent process. The uptake of this compound is also temperature-dependent, with a Km of 1.6 X 10(-4)M for the high-affinity system. D-alpha-Aminoadipate has characteristics similar to those displayed by the L-isomer but to a lesser degree. L-Glutamate and L-aspartate inhibit the uptake of L-alpha-aminoadipate. D- and L-alpha-Aminoadipate are, respectively, weak uncompetitive and weak competitive inhibitors for the uptake of L-glutamate and L-aspartate. Both enantiomers inhibit GABA uptake but in quite different ways. The release of L-alpha-aminoadipate from the cerebral cortical slices is stimulated by a high concentration of K+ ions in the presence of Ca2+ in the perfusion buffer; the D-isomer displays this property to a lesser degree. The omission of Ca2+ markedly reduces the release of these two compounds. Less than 10% of the preloaded D- and L-alpha-aminoadipate are metabolized by the cerebral cortex during 40 min of superfusion. The possibility of L-alpha-aminoadipate as a neurotransmitter candidate is discussed.

2-Aminoadipic Acid↗
PubMed1980

A simple method for the preparation of D-alpha-aminoadipic acid.

High quantity (1 g and more) of racemically and chromatographically pure D-alpha-aminoadipic acid was prepared by selective metabolism of the L-isomer of the commercially available DL-alpha-aminoadipate by Pseudomonas putida. The overall yield of this preparation averaged 40%. The final product has [a]25D value of -25 degrees. This procedure can be useful in the synthesis of high purity D-alpha-amino-adipate, a compound shown recently to be a useful tool in the study of neurotransmission mechanism mediating synaptic excitation.

2-Aminoadipic Acid↗
PubMed1978

Aspartokinase of Streptococcus mutans: purification, properties, and regulation.

Aspartokinase from Streptococcus mutans BHT was purified to homogeneity and characterized. The molecular weight of the native enzyme was estimated to be 242,000 by gel filtration. Cross-linking of aspartokinase with dimethyl suberimidate and polyacrylamide gel electrophoresis of the amidinated enzyme in the presence of sodium dodecyl sulfate showed the enzyme to be composed of six identical subunits with a molecular wieght of 40,000. The optimal pH range for enzyme activity was 6.5 to 8.5. The apparent Michaelis-Menten constants for aspartate and ATP were 5.5 and 2.2 mM, respectively. The enzyme was stable within the temperature range of 10 to 35 degrees C. Aspartokinase was not feedback inhibited by individual amino acids, but was concertedly inhibited by L-lysine and L-threonine (93.5% inhibition at 10 mM each). The inhibition was noncompetitive with respect to aspartate (Ki = 10 mM) and mixed with respect to ATP. L-Threonine methyl ester and L-threonine amide were able to substitute for L-threonine in feedback inhibition, but the requirement for L-lysine uas strict. The feedback inhibitor pair protected the enzyme against heat denaturation. Aspartokinase synthesis was repressed by L-threonine; this repression was enhanced by L-lysine, but was slightly attenuated by L-methionine.

Adenosine Triphosphate↗
PubMed1977

Glutarate semialdehyde dehydrogenase of Pseudomonas. Purification, properties, and relation to L-lysine catabolism.

The lysine-induced glutarate semialdehyde dehydrogenase of Pseudomonas was purified to electrophoretic homogeneity from a mutant strain lacking delta-aminovalerate transaminase. The properties of the enzyme, including molecular weight, amino acid composition, electrophoretic behavior, and kinetic features, distinguish it from similar dehydrogenases induced in the same cell strain by hydroxyproline or by glucarate. Enzyme induction patterns and the growth behavior of a mutant deficient in glutarate semialdehyde dehydrogenase clearly relate this enzyme to the so-called delta-aminovalerate pathway of L-lysine catabolism. Induction studies also indicate that delta-aminovalerate is a better inducer of the dehydrogenase than L-lysine. Cells of a mutant strain lacking delta-aminovalerate transaminase contained higher levels of the dehydrogenase, presumably as a result of the accumulation of delta-aminovalerate, making this mutant a useful preparative source of the enzyme. The marked reduction of lysine-inducible glutarate semialdehyde dehydrogenase in a mutant strain permitted assessment of the basal levels of hydroxyproline/glucarate-inducible ketoglutarate semialdehyde dehydrogenases not possible in wild type cells.

Aldehyde Oxidoreductases↗
PubMed1974

D-lysine catabolic pathway in Pseudomonas putida: interrelations with L-lysine catabolism.

The isolation of several mutant strains blocked in l-lysine degradation has permitted an assessment of the physiological significance of enzymatic reactions related to lysine metabolism in Pseudomonas putida. Additional studies with intact cells involved labeling of metabolic intermediates from radioactive l- or d-lysine, and patterns of enzyme induction in both wild-type and mutant strains. These studies lead to the conclusions that from l-lysine, the obligatory pathway is via delta-aminovaleramide, delta-aminovalerate, glutaric semialdehyde, and glutarate, and that no alternative pathways from l-lysine exist in our strain. A distinct pathway from d-lysine proceeds via Delta(1)-piperideine-2-carboxylate, l-pipecolate, and Delta(1)-piperideine-6-carboxylate (alpha-aminoadipic semialdehyde). The two pathways are independent in the sense that certain mutants, unable to grow on l-lysine, grow at wild-type rates of d-lysine, utilizing the same intermediates as the wild type, as inferred from labeling studies. This finding implies that lysine racemase in our strain, while detectable in cell extracts, is not physiologically functional in intact cells at a rate that would permit growth of mutants blocked in the l-lysine pathway. Pipecolate oxidase, a d-lysine-related enzyme, is induced by d-lysine and less efficiently by l-lysine. Aminooxyacetate virtually abolishes the inducing activity of l-lysine for this enzyme, suggesting that lysine racemase, although functionally inactive for growth purposes, may still have regulatory significance in permitting cross-induction of d-lysine-related enzymes by l-lysine, and vice versa. This finding suggests a mechanism in bacteria for maintaining regulatory patterns in pathways that may have lost their capacity to support growth. In addition, enzymatic studies are reported which implicate Delta(1)-piperideine-2-carboxylate reductase as an early step in the d-lysine pathway.

Acetates↗
PubMed1970

D-glucaric acid and galactaric acid catabolism by Agrobacterium tumefaciens.

Cell-free extract (crude extract) of Agrobacterium tumefaciens grown on d-glucuronate or d-glucarate converts d-glucarate and galactarate to a mixture of 2-keto-3-deoxy- and 4-deoxy-5-keto-d-glucarate. These compounds are then converted by partially purified crude extract to an intermediate tentatively identified as 2,5-diketoadipate. The same enzyme preparation further decarboxylates this intermediate to alpha-ketoglutarate semialdehyde, which is subsequently oxidized in a nicotinamide adenine dinucleotide-dependent reaction to alpha-ketoglutaric acid. Since A. tumefaciens converts d-glucuronic acid to d-glucarate, a pathway from d-glucuronate to alpha-ketoglutarate in A. tumefaciens was determined.

Adipates↗