Oxidative deamination of S-(1-carboxyethyl)-L-cysteine and S-(1-carboxypropyl)-L-cysteine by L-aminoacid oxidase

S-(1-carboxyethyl)-L-cysteine (1-CEC) and S-(1-carboxypropyl)-L-cysteine (1-CPC) are oxidatively deaminated by L-aminoacid oxidase with consumption of half a mole of oxygen per mole of substrate in the presence of catalase. This reaction gives rise to the corresponding alpha-ketoacids, identified by some chemical and chromatographic tests and by comparison with synthetic compounds. It has been possible, therefore, to demonstrate that S-(1-carboxyethyl)-thiopvruvic acid (1-CETP) and S-(1-carboxypropyl)-thiopvruvic acid (1-CPTP) are the main products of oxidative deamination of 1-CEC and 1-CPC.     

Oxidative deamination of Se-(1-carboxyethyl)-,Se-(1-carboxypropyl)- and Se-(2-carboxyethyl)-selenocysteine by snake venom L-aminoacid oxidase

Details are reported for the synthesis of Se-(1-carboxyethyl)-selenocysteine (1-CESeC), Se-(1-carboxypropyl)-selenocysteine (1-CPSeC) and Se-(2-carboxyethyl)-selenocysteine (2-CESeC). They can be obtained in pure cristalline form with good yield. Some chromatographic properties, useful for their identification, are described. The three aminoacids are good substrates for snake venom L-aminoacid oxidase, giving the corresponding alpha-ketoacids as reaction products.     

UV-Photolysis of S-(cis-l-Propenyl)-L-cysteine in Oxygen-free Aqueous Solution

From the comparisons of mass spectral fragmentations and gas chromatographic retention times with reference compounds, volatile flavor products from UV-photolysis of S-(cis-l-propenyl)-L-cysteine in oxygen-free aqueous solutions were identical with propanal, 2-methyl-2-pentenal, n-propyl mercaptan, allyl mercaptan, 1-propenyl mercaptan, 2-methylthiophene, 3-methylthiophene, 2,3-dimethylthiophene, 2,4-dimethylthiophene, 2,5-dimethylthiophene, 3,4-dimethylthiophene, n-propyl 1-propenyl sulfide and di- 1-propenyl sulfide. Moreover, from the comparison of two-dimentional thin-layer chromatograms with reference compounds, ninhydrin-positive products with interest in terms of the degradation mechanism were identical with alanine and cystine. It seems that above-mentioned thiophene derivatives are produced via 1-propenyl thiyl radicals.     

Cytotoxicity of S-(1,2-dichlorovinyl)glutathione and S-(1,2-dichlorovinyl)-L-cysteine in isolated rat kidney cells

S-(1,2-Dichlorovinyl)glutathione (DCVG) and S-(1,2-dichlorovinyl)-L-cysteine (DCVC) produced time- and concentration-dependent cell death in isolated rat kidney proximal tubular cells. AT-125 blocked and glycylglycine potentiated DCVG toxicity, indicating that metabolism by gamma-glutamyltransferase is required. S-(1,2-Dichlorovinyl)-L-cysteinylglycine, a putative metabolite of DCVG, also produced cell death, which was prevented by 1,10-phenanthroline, phenylalanylglycine, and aminooxyacetic acid, inhibitors of aminopeptidase M, cysteinylglycine dipeptidase, and cysteine conjugate beta-lyase, respectively. Aminooxyacetic acid and probenecid protected against DCVC toxicity, indicating a role for metabolism by cysteine conjugate beta-lyase and organic anion transport, respectively. DCVC produced a small decrease in cellular glutathione concentrations and did not change cellular glutathione disulfide concentrations or initiate lipid peroxidation. DCVC caused a large decrease in cellular glutamate and ATP concentrations with a parallel decrease in the total adenine nucleotide pool; these changes were partially prevented by aminooxyacetic acid. Both DCVG and DCVC inhibited succinate-dependent oxygen consumption, but DCVC had no effect when glutamate + malate or ascorbate + N,N,N',N'-tetramethyl-p-phenylenediamine were the electron donors. DCVC inhibited mitochondrial, but not microsomal, Ca2+ sequestration. These alterations in mitochondrial function were partially prevented by inhibition of DCVG and DCVC metabolism and were strongly correlated with decreases in cell viability, indicating that mitochondria may be the primary targets of nephrotoxic cysteine S-conjugates.     

芦笋抗S—（2—氨乙基）—L—半胱氨酸（AEC）变异体的筛选

S-(2-氨乙基)-L-半胱氨酸(AEC)可抑制芦笋愈伤组织的生长,此抑制作用可被赖氨酸或甲硫氨酸部分解除。用0.5mmol/L的AEC进行筛选,得到抗性愈伤组织AR10并再生植株。AR10愈伤组织经一年多的继代培养,在离开选择剂组培继代两代后仍保持对AEC的抗性。抗性系愈伤组织还表现出对2mmol/L的半胱氨酸具交叉抗性,对1mmol/L的赖氨酸加苏氨酸表现部分交叉抗性。AR10再生植株一部分保持对AEC的抗性,而一部分则无抗性。对抗性愈伤组织及其再生植株的氨基酸分析表明,愈伤组织内游离赖氨酸、苏氨酸、甲硫氨酸都有增加,而在再生植株内却发现半胱氨酸和赖氨酸的特异性增加,分别是对照植株的5.4和4.6倍。     

Modification of polynucleotides by a fragment produced by enzymatic cleavage of S-(1, 2-dichlorovinyl)-L-cysteine

Polyribo- and polydeoxyribonucleotides were allowed to react with ³⁵S-(1,2-dichlorovinyl)-L-cysteine (DCVC) in presence of a bovine kidney lyase yielding products which were substituted to varying degrees with an alkylating thiovinyl fragment (AF) released from DCVC. Polydeoxyribonucleotides were more extensively substituted than polyribonucleotides. Double stranded homopolymer pairs were much less effective as acceptors of (AF) than single stranded polymers. Nucleotide substitution occurred only at the polymer level. Enzymatic hydrolysis of (AF)-substituted polymers yielded dinucleotides which contained an (AF) fragment apparently covalently linked in unknown fashion. (AF)-substituted polynucleotides had reduced ability to form helical complexes with complementary polynucleotides, as revealed by hypochromicity, melting transition and renaturation.     

Synthesis of [2-3H-ethyl]S-(2-chloro-1,1,2-trifluoroethyl)-L-cysteine and its use in covalent-binding studies.

Metabolism of S-(2-chloro-1,1,2-trifluoroethyl)-L-cysteine (CTFC) yields chlorofluorothioacetyl fluoride, which reacts with cellular proteins to form stable lysine adducts. Little is known about the subcellular localization of these protein adducts or about their role in CTFC-induced nephrotoxicity. A method for the synthesis of CTFC and other cysteine S-conjugates labeled with 3H at the S-alkyl or S-alkenyl position would be useful in studies of S-conjugate metabolism and toxicity. Reaction of L-cysteine, chlorotrifluoroethene, 1,8-diazabicyclo[5.4.0]undec-7-ene, and 3H-labeled water followed by repeated crystallization yielded radiochemically pure [3H]CTFC (235 mg, 20% yield; sp act 1.07 x 10(9) Bq/mmol), which was identical to CTFC by TLC, 1H NMR, and 19F NMR. 3H NMR revealed a doublet of triplets at 6.5 ppm with geminal and vicinal T-F couplings of 51.5 and 6.0 Hz, respectively, consistent with the proposed structure. When 2H-labeled water was used, [2H]CTFC was formed, and its structure was confirmed by 1H and 19F NMR, FAB-MS, and TLC. Analysis of renal and hepatic subcellular fractions of rats given 1, 10, or 100 mumol/kg [3H]CTFC showed a dose-dependent binding of 3H-containing metabolites to liver and kidney proteins.