Inhibition of Glutamate Dehydrogenase in Brain Mitochondria and Synaptosomes by Mg2+ and Polyamines: A Possible Cause for Its Low In Vivo Activity

Abstract: Magnesium and the polyamines putrescine, spermidine, and spermine inhibited the activity of glutamate dehydrogenase in permeabilized rat brain mitochondria in a concentration-dependent manner. The inhibitory effect was observed on both the reductive amination of 2-oxoglutarate and oxidative deamination of glutamate, as well as in the presence and absence of ADP and leucine, the allosteric activators of the enzyme. Kinetic studies at various concentrations of substrates showed that inhibition by magnesium and spermine was very pronounced at 2-oxoglutarate concentrations less than 0.5 m M and NADH levels less than 0.08 m M . The presence of the former compounds also accentuated the inhibitory effect of high concentrations of 2-oxoglutarate (>2.0 m M ) and NADH (>0.32 m M ). Addition of magnesium and spermine to suspensions of synaptosomes decreased the amount of ammonia produced from glutamate. It is suggested that polyamines and magnesium, normal constituents of mammalian brain, are responsible, at least in part, for the low glutamate dehydrogenase activity in vivo.     

Modulation by angiotensin III of nociception-related and arterial pressure-related neuronal responsiveness in the nucleus reticularis gigantocellularis of the rat

We evaluated possible modulation by angiotensin III (AIII) of the interactive effect of noxious stimuli and elevation in systemic arterial pressure on the responsiveness of neurons in the nucleus reticularis gigantocellularis (NRGC) of the medulla oblongata. Combined extracellular single-neuron recording and microiontophoresis were carried out on male, adult Sprague-Dawley rats anesthetized with pentobarbital sodium. The responsiveness of NRGC neurons to nociception (tail clamp) and/or transient hypertension elicited by phenylephrine (5 μg/kg, i.v.), in the absence or presence of AIII, was used as the experimental index. Microiontophoretic application of the heptapeptide suppressed the responses of spontaneously active NRGC neurons to individually delivered nociception or hypertension. Interestingly, the preferential reduction in responsiveness to tail clamp upon simultaneous elevation in arterial pressure was reversed to one that favored nociception in the presence of AIII. These actions of the heptapeptide appeared to be receptor-specific, since they were discernibly blocked by its selective antagonist, Ile⁷-angiotensin III. Our results reveal that neuropeptides such as AIII may differentially modulate neuronal responsiveness according to the prevailing physiologic input(s) to the central nervous system of the animal.     

UDP-glucose dehydrogenase from bovine liver: primary structure and relationship to other dehydrogenases.

The primary structure of bovine liver UDP-glucose dehydrogenase (UDPGDH), a hexameric, NAD(+)-linked enzyme, has been determined at the protein level. The 52-kDa subunits are composed of 468 amino acid residues, with a free N-terminus and a Ser/Asn microhetergeneity at one position. The sequence shares 29.6% positional identity with GDP-mannose dehydrogenase from Pseudomonas, confirming a similarity earlier noted between active site peptides. This degree of similarity is comparable to the 31.1% identity vs. the UDPGDH from type A Streptococcus. Database searching also revealed similarities to a hypothetical sequence from Salmonella typhimurium and to "UDP-N-acetyl-mannosaminuronic acid dehydrogenase" from Escherichia coli. Pairwise identities between bovine UDPGDH and each of these sequences were all in the range of approximately 26-34%. Multiple alignment of all 5 sequences indicates common ancestry for these 4-electron-transferring enzymes. There are 27 strictly conserved residues, including a cysteine residue at position 275, earlier identified by chemical modification as the expected catalytic residue of the second half-reaction (conversion of UDP-aldehydoglucose to UDP-glucuronic acid), and 2 lysine residues, at positions 219 and 338, one of which may be the expected catalytic residue for the first half-reaction (conversion of UDP-glucose to UDP-aldehydoglucose). A GXGXXG pattern characteristic of the coenzyme-binding fold is found at positions 11-16, close to the N-terminus as with "short-chain" alcohol dehydrogenases.(ABSTRACT TRUNCATED AT 250 WORDS)     

广东种子植物区系地理成分研究

广东种子植物区系共有２１９科１４３４属４９８６种。其中：①热带区系成分占有较大的比例,约占非世界属的６２．３％,表明广东区系受到热带区系的强烈渗透,次之为亚热带（至亚热带山地）成分约占３２．２％,及中国特有分布占５．６％;②以单型属（７２属）,单种属（５８５属）和寡种属（５６２属）占绝对优势,共占８５．０％,充分显示了广东区系的古老性和过渡性特点;③从区系组成和植被结构的特征成分来看,以华夏植物区系成分为主,包括：亚热带分布,亚热带山地分布,中国特有分布,东亚—北美间断分布,及一些亚洲热带分布中以亚热带为主的属：④在这些属中有东亚特有属１３２属,中国特有属７５属以及华南—西南—中南半岛特有属６８属等;这些属的存在毫无疑问地说明,广东区系是华夏植物区系的核心地区之一,同时也显示了热带区系与亚热带区系有着不可分隔的统一性。     

杉木连栽土壤对其幼树生长的影响

在会同林区采用３种不同连栽杉木次数的土壤,进行杉木幼树盆栽试验．结果表明,杉木连栽不利其幼树生长．连裁使幼树高生长下降３７—４０％,基径下降１９—２８％,冠幅下降２１—２９％,总生物量减少４５—５０％．杉木连栽还造成土壤养分递减,即腐殖质减少７—２８％,速效养分下降２３—２８％,土壤微生物数量和活性也大大降低．因此,杉木纯林连栽经营方式应尽快改变．     

Control of gluconeogenic growth by pps and pck in Escherichia coli.

It is well-known that Escherichia coli grows more slowly on gluconeogenic carbon sources than on glucose. This phenomenon has been attributed to either energy or monomer limitation. To investigate this problem further, we varied the expression levels of pck, encoding phosphoenolpyruvate carboxykinase (Pck), and pps, encoding phosphoenolpyruvate synthase (Pps). We found that the growth rates of E. coli in minimal medium supplemented with succinate and with pyruvate are limited by the levels of Pck and Pps, respectively. Optimal overexpression of pck or pps increases the unrestricted growth rates on succinate and on pyruvate, respectively, to the same level attained by the wild-type growth rate on glycerol. Since Pps is needed to supply precursors for biosyntheses, we conclude that E. coli growing on pyruvate is limited by monomer supply. However, because pck is required both for biosyntheses and catabolism for cells growing on succinate, it is possible that growth on succinate is limited by both monomer and energy supplies. The growth yield with respect to oxygen remains approximately constant, even though the overproduction of these enzymes enhances gluconeogenic growth. It appears that the constant yield for oxygen is characteristic of efficient growth on a particular substrate and that the yield is already optimal for wild-type strains. Further increases in either Pck or Pps above the optimal levels become growth inhibitory, and the growth yield for oxygen is reduced, indicating less efficient growth.     

Relationship between receptor/ligand binding affinity and adhesion strength.

Receptor-mediated cell adhesion is a central phenomenon in many physiological and biotechnological processes. Mechanical strength of adhesion is generally presumed to be related to chemical affinity of receptor/ligand bonds, but no experimental study has been previously directed toward this issue. Here we investigate the dependence of receptor/ligand adhesion strength on bond affinity using a radial fluid flow chamber assay to measure the force needed to detach polystyrene beads covalently coated with immunoglobulin G from glass surfaces covalently coated with protein A. A spectrum of animal species sources for immunoglobulin G permits examination of three decades of protein A/immunoglobulin G binding affinity. Our results for this model system demonstrate that adhesion strength varies with the logarithm of the binding affinity, consistent with a prediction from the theoretical model by Dembo et al. (Dembo, M., D.C. Torney, K. Saxman, and D. Hammer. 1988. Proc. R. Soc. Lond. Ser. B 234:55-83).     

复合四倍体异育银鲫个体间遗传异质性的研究

用聚丙烯酰胺梯度凝胶电泳,分析比较了４个不同的银鲫雌核发育系,红鲤和复合四倍体异育银鲫鳍的５种不同的同工酶及蛋白表型的差异,表明复合四倍体异育银鲫表型上的差异主要是来自银鲫种内的遗传差异（不同的雌核发育系）。来源于同一个银鲫雌核发育系的复合四倍体异育银鲫,个体间的ＥＳＴ同工酶出现差异,并与父体红鲤的ＥＳＴ同工酶的多态性相关,因此,复合四倍体异育银鲫个体间的异质性也包含了来自父本的遗传影响。在所检测