Involvement of D-amino acid oxidase in elimination of D-serine in mouse brain

The physiological role of D-amino acid oxidase (EC 1. 4. 3. 3) in mouse brain is described. The presence of D-enantiomers of neutral common amino acids was surveyed in the brain. D-serine was shown to be present at high concentration only in regions where the enzyme activity was low. In normal mice whose D-amino acid oxidase activity was much higher in the cerebellum than in the cerebrum, free D-serine content was apparently lower in the cerebellum than in the cerebrum. In mice of a mutant strain lacking D-amino acid-oxidase activity, the free D-serine level was remarkably high both in the cerebrum and cerebellum. The results suggest that the enzyme is involved in the elimination of free D-serine in the cerebellum.     

L-谷氨酸氧化酶高产菌株的快速筛选及诱变育种

用谷氨酸氧化酶测试液浸湿白卷纸置于菌落上,根据白卷纸上显示红色快慢、深浅,就能大致测得该菌落菌株产酶能力高低。该法可靠性大、工作量小、操作简便,显示结果快。利用该法,配合亚硝酸诱变,在短暂三个月内,将L-谷氨酸氧化酶产生菌株产酶能力提高了约25倍。     

固定化条件对葡萄糖酶电极响应性能的影响

用微铂盘电极和交联法制得的酶膜构成葡萄糖酶电极，测定其电流响应特性，研究了酶的固定化条件：酶含量以及载体蛋白和交联剂的用量等，对响应特性的影响。提出较适宜的酶电极制备方法，并检测了所得的电极在不同ｐＨ溶液中的灵敏度，指出其最佳操作条件。     

异羟肟酸比色法测定电解混合液中葡萄糖酸的含量

本文报导用异羟肟酸比色法,分析葡萄糖电解氧化制备葡萄糖酸盐过程的电解混合液中葡萄糖酸根含量的研究结果,文中对实验条件及影响因素进行了探讨。实验证明,本方法具有分析方法简单、分析数据稳定、重现性好、可靠、方便等特点。     

Administration of the antitumor drug mitoguazone protects normal thymocytes against spontaneous and etoposide-induced apoptosis

The suggestion has been made that polyamines may be involved in the control of cell death, since exceedingly high or low levels induce apoptosis in different cell systems. For a deeper insight into the relationship between apoptosis and polyamine metabolism, we investigated in vitro the effect on rat thymocytes of mitoguazone (MGBG, which inhibits S-adenosylmethionine decarboxylase, i.e. a key enzyme in the polyamine biosynthetic pathway). Thymocytes were selected as an especially suitable model system, since they undergo spontaneous apoptosis in vivo and can be easily induced to apoptose in vitro by etoposide, used here as an apoptogenic agent. MGBG protected thymocytes from both spontaneous and drug-induced apoptosis, and this protective effect was associated with a decrease in polyamine oxidase activity and total polyamine levels.Received 7 July 2004; received after revision 2 September 2004; accepted 9 September 2004     

The superoxide-generating NADPH oxidase: structural aspects and activation mechanism

Flavocytochrome b ₅₅₈ is the catalytic core of the respiratory-burst oxidase, an enzyme complex that catalyzes the NADPH-dependent reduction of O₂ into the superoxide anion O₂ ^- in phagocytic cells. Flavocytochrome b ₅₅₈ is anchored in the plasma membrane. It is a heterodimer that consists of a large glycoprotein gp91phox (phox for phagocyte oxidase) (β subunit) and a small protein p22phox (α subunit). The other components of the respiratory-burst oxidase are water-soluble proteins of cytosolic origin, namely p67phox, p47phox, p40phox and Rac. Upon cell stimulation, they assemble with the membrane-bound flavocytochrome b ₅₅₈ which becomes activated and generates O₂ ^-. A defect in any of the genes encoding gp91phox, p22phox, p67phox or p47phox results in chronic granulomatous disease, a genetic disorder characterized by severe and recurrent infections, illustrating the role of O₂ ^- and the derived metabolites H₂O₂ and HOCl in host defense against invading microorganisms. The electron carriers, FAD and hemes b, and the binding site for NADPH are confined to the gp91phox subunit of flavocytochrome b ₅₅₈ . The p22phox subunit serves as a docking site for the cytosolic phox proteins. This review provides an overview of current knowledge on the structural organization of the O₂ ^--generating flavocytochrome b ₅₅₈ , its kinetics, its mechanism of activation and the regulation of its biosynthesis. Homologues of gp91phox, called Nox and Duox, are present in a large variety of non-phagocytic cells. They exhibit modest O₂ ^--generating oxidase activity, and some act as proton channels. Their role in various aspects of signal transduction is currently under investigation and is briefly discussed. Received 28 May 2002; received after revision 20 June 2002; accepted 24 June 2002     

Lipid interactions with transmembrane proteins

Magnetic resonance results, principally from ²H-nuclear magnetic resonance, indicate that the mean lipid-chain ordering at the surface of transmembrane proteins is comparable to that in fluid lipid bilayers. Principally, it is the requirement for matching the hydrophobic lengths of lipid and protein that modulates the degree of chain ordering at the lipid-protein interface. The distribution of chain order parameters is, nonetheless, broader in the presence of integral proteins than in fluid lipid bilayers. The chain configurations of the phospholipids that are resolved in crystals of integral membrane proteins display considerable conformational heterogeneity. Chain C–C dihedral angles are, however, not restricted to the energetically allowable trans and gauche rotamers. This indicates that the chains of a given lipid do not have a unique configuration in protein crystals.     

Assignment of Soret MLCT band of reduced form of copper binuclear cluster in cytochrome c oxidase film

Low concentration of dithionite results in the reduction of Cu-Cn binuclear and heine a active sites of the cytochrome c oxidase thin solid film immersed in the acidic phosphate buffer, but Fe-Cu binuclear center keeps in the oxidation state. It manifests as a negative peak at 426 nm and a positive one at -408 nln in the difference spectra induced by dithionite. The former implies decrease of the oxidized form of heme a center, that is, Fea^3 →Fea^2 . And the latter results from the contribution of metal-ligand charge transfer (MLCT) transition in the reduced binuclear Cu-Cu cluster, rather than from that of heine a center. This stronger Soret MLCT band must be helpful to overcoming the difficulty in distinguishing the weaker copper sign from the stronger one of iron when studying copper-iron protein.     

‘Classical’ and ‘new’ diabetogens—comparison of their effects on isolated rat pancreatic islets in vitro

This study compares functional and morphological alterations caused by application of alloxan, streptozotocin, xanthine oxidase/hypoxanthine (generation of reactive oxygen species), or S-nitroso-N-acetyl-D,L-penicillamine (SNAP, liberation of nitric oxide) to isolated rat pancreatic islets in vitro. In perifusion experiments, membrane leakage—detected by non-stimulated insulin release—was found after application of all drugs, but showed a substance-specific time pattern. Twenty-four hours after application of the classical diabetogens (alloxan or streptozotocin), potassium chloride- and glucose-stimulated insulin secretion were markedly reduced, while a persistent reduction was observed neither after exposure to xanthine oxidase/hypoxanthine, nor to SNAP. Morphological analysis of the islets revealed that nearly all β-cells were destroyed following alloxan or streptozotocin treatment, while the majority of β-cells were configured regularly after application of xanthine oxidase/hypoxanthine or SNAP. Necrotic cells found after xanthine oxidase/hypoxanthine usually differed in morphology from those observed after application of the classical diabetogens. While the former cells were characterised by swollen nuclei, the latter had shrunken nuclei with irregular condensed chromatin. Apoptosis was found only following nitric oxide exposure. Due to these differences, it seems unlikely that alloxan, streptozotocin, xanthine oxidase/hypoxanthine, and nitric oxide have a common major feature in their toxic action. Received 16 September 1999; received after revision 15 November 1999; accepted 26 November 1999     

基于铅笔芯和多巴胺电聚合膜固定葡萄糖氧化酶的高灵敏酶电极

市售8B铅笔芯经镀铂处理后,将葡萄糖氧化酶共沉积到新型多巴胺电氧化聚合膜中,最外层再修饰聚邻氨基酚薄膜,研制出新型葡萄糖氧化酶修饰电极用于葡萄糖检测.