Insights into acylphosphatase structure and catalytic mechanism

Acylphosphatase is one of the smallest enzymes known (about 98 amino acid residues). It is present in organs and tissues of vertebrate species as two isoenzymes sharing over 55% of sequence homology; these appear highly conserved in differing species. The two isoenzymes can be involved in a number of physiological processes, though their effective biological function is not still certain. The solution and crystal structures of different isoenzymes are known, revealing a close packed protein with a fold similar to that shown by other phosphate-bind ing proteins. The structural data, together with an extended site-directed mutagenesis investigation, led to the identification of the residues involved in enzyme catalysis. However, it appears unlikely that these residues are able to perform the full catalytic cycle: a substrate-assisted catalytic mechanism has therefore been proposed, in which the phosphate moiety of the substrate could act as a nucleophile activating the catalytic water molecule. Received 12 November 1996; accepted 27 November 1996     

Thyroxine-induced redistribution of creatine kinase isoenzymes in rat cardiomyocyte cultures

Summary The present study demonstrates a change occurring in the creatine-kinase isoenzyme profile of cardiomyocyte cultures induced by a chronic administration of excessive amounts of thyroid hormones (TH). This change is manifested by an increased level of the CK-BB isoenzyme, generally at the expense of CK-MM isoenzyme. The elevation of CK-BB is probably a result of a specific effect of TH through activation of gene expression, rather than a contribution of an increased number of non-myocardial cells. The implications of these results in the diagnosis of heart failures are discussed.     

福建山麻鸭LDH同工酶组织特异性的研究

采用聚丙烯酰胺等电聚焦电泳方法对福建山麻鸭的心、肝、肺、肾及血清５种器官组织的乳酸脱氢酶同工酶进行分离，结果得到１４－１７条电泳区带，其等电点范围在ＰＨ４．８－９．１之间，多数集居中性区，各器官组织电泳图谱表现出明显的正极端的不均一性和特有的活性含量的分布差异，这些资料对福建家鸭种质资源调查和开发利用颇为用。     

猪心肌乳酸脱氢酶的提纯工艺及酶反应最适条件

研究了适合于工业化生产的猪心肌乳酸脱氢酶的初步提取工艺，经硫酸铵分级沉淀得到酶的比活为86．5U／mg，再经DEAE离子交换层析比活可达525．0U／mg，酶活总收率为54．5％．并对酶促反应的最适条件进行了研究，其最适反应pH为7．6～7．8，最适反应底物浓度分别为NADH5mg／mL和丙酮酸钠25mmol／L。     

Molecular mechanism for the production of multiple form of MM creatine kinase

Summary Incubation of human, canine or rabbit MM creatine kinase with carboxypeptidase-N or B resulted in the production of 2 additional enzyme forms with increased anodal migration on polyacrylamide gels. The C-terminal amino acid of tissue MM creatine kinase from all 3 species was shown to be lysine, a specific substrate for carboxypeptidase-N and B.     

Lactate dehydrogenase and glutamate dehydrogenase activities in the circumventricular organs of rat brain following neonatal monosodium glutamate

Glutamate (glu) an excitatory neurotransmitter amino acid, is present in high concentrations in the mammalian central nervous system and is the most abundant amino acid in our daily diet. In the present study the activities of lactate dehydrogenase (LDH) and glutamate dehydrogenase (GDH) were evaluated in the circumventricular organs (CVO) of the brain in 25-day-old rats following MSG administration at a dose of 4 mg/g b.wt during the first ten days of life. The results show the LDH activity increased to 265% of that in the control (p<0.001), whereas GDH activity was significantly decreased (p<0.05), The great elevation in LDH, a cytoplasmic marker enzyme, is apparently due to cytoskeletal changes brought about as a consequence of glu toxicity, whereas lowered GDH activity indicates altered glu homostasis in the blood-brain-barrier deficient areas following neonatal exposure to glu.     

虫草素和虫草素/双层氢氧化物纳米复合物对HeLa细胞生长抑制

为解决虫草素在体内被腺苷脱氢酶(ADA)快速降解的问题,制备虫草素/LDH复合物,研究其在体内外对虫草素的缓释以及对HeLa细胞的生长抑制作用.结果表明:当虫草素/LDH复合物中虫草素质量浓度为30,60,90μg·mL-1时,复合物对HeLa细胞的生长抑制率分别为52.4%、61.3%、79.6%,与虫草素对照组比较,P<0.01.显微镜观察发现,当虫草素质量浓度60μg·mL-1时,虫草素/LDH复合物使大部分HeLa细胞死亡;虫草素的对照组HeLa细胞死亡较少,但细胞内部明显出现空泡.虫草素/LDH复合物体外释放表明,虫草素插入LDH双层结构之后,使虫草素释放速度明显被降低.对SD大鼠腹腔给药后,虫草素/LDH复合物血药最高质量浓度为82.1μg·mL-1,在药后1.5h出现,虫草素对照组的血药最高质量浓度为75.3μg·mL-1,在给药后1h出现.     

小白鼠部分器官中乳酸脱氢酶（LDH）分布特异性的探讨

本文证实了乳酸脱氢酶（LDH）在小白鼠部分器官中的分布特异性.指出了乳酸脱氢酶五种同工酶在小白鼠的心肌、骨骼肌、肾脏、脑和肝脏中均存在，并且在心肌、脑和肾脏中LDH-1（H4）占优势;而在骨骼肌和肝脏中以LDH-5（M4）占优势;分别与各自的生理功能相适应.     

油菜花序轴组织培养中愈伤组织同工酶和可溶性蛋白质变化的研究

本文报道了在愈伤组织形成过程中,酯酶、过氧化物酶同工酶及可溶性蛋白质含量的变化,并讨论了愈伤组织形成与这些大分子变化之间的关系。