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801.
F. H. Wilcox 《Biochemical genetics》1973,10(1):69-78
Two strains of mice were shown to differ in relative proportion of two major protein bands of liver under acid conditions of electrophoresis. Genetic control was autosomal and by a pair of dominant and recessive genes. The difference was observed only if liver homogenates were extracted by treating with dilute acid and alkali. The two protein bands were identified as albumin on the basis of electrophoretic migration, molecular weight, and immunological response. No differences were found between strains in relative amounts of comparable bands of plasma. However, in studies with extraction of mixtures, liver homogenates of one strain but not another were capable of converting a large proportion of albumin from several sources to a faster-migrating form. It was concluded that the strains differ in a factor in liver capable of causing a secondary modification of the albumin molecule. 相似文献
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Elizabeth Wilcox Andrei V. Blokhin Ernest Hamel 《Archives of biochemistry and biophysics》2009,484(1):55-62
GDP inhibits paclitaxel-induced tubulin assembly without GTP when the tubulin bears GDP in the exchangeable site (E-site). Initially, we thought inhibition was mediated through the E-site, since small amounts of GTP or Mg2+, which favors GTP binding to the E-site, reduced inhibition by GDP. We thought trace GTP released from the nonexchangeable site (N-site) by tubulin denaturation was required for polymer nucleation, but microtubule length was unaffected by GDP. Further, enhancing polymer nucleation reduced inhibition by GDP. Other mechanisms involving the E-site were eliminated experimentally. Upon finding that ATP weakly inhibited paclitaxel-induced assembly, we concluded that another ligand binding site was responsible for these inhibitory effects, and we found that GDP was not binding at the taxoid, colchicine, or vinca sites. There may therefore be a lower affinity site on tubulin to which GDP can bind distinct from the E- and N-sites, possibly on α-tubulin, based on molecular modeling studies. 相似文献
807.
Nicholas E. Grossoehme Anne M. Spuches Dean E. Wilcox 《Journal of biological inorganic chemistry》2010,15(8):1183-1191
The thermodynamics of metals ions binding to proteins and other biological molecules can be measured with isothermal titration
calorimetry (ITC), which quantifies the binding enthalpy (ΔH°) and generates a binding isotherm. A fit of the isotherm provides the binding constant (K), thereby allowing the free energy (ΔG°) and ultimately the entropy (ΔS°) of binding to be determined. The temperature dependence of ΔH° can then provide the change in heat capacity (ΔC
p°) upon binding. However, ITC measurements of metal binding can be compromised by undesired reactions (e.g., precipitation,
hydrolysis, and redox), and generally involve competing equilibria with the buffer and protons, which contribute to the experimental
values (K
ITC, ΔH
ITC). Guidelines and factors that need to be considered for ITC measurements involving metal ions are outlined. A general analysis
of the experimental ITC values that accounts for the contributions of metal–buffer speciation and proton competition and provides
condition-independent thermodynamic values (K, ΔH°) for metal binding is developed and validated. 相似文献
808.
Marshall B. Elam George S. Cowan Jr Robert J. Rooney M. Lloyd Hiler Chandrahasa R. Yellaturu Xiong Deng George E. Howell Edwards A. Park Ivan C. Gerling Divyan Patel J. Christopher Corton Lauren M. Cagen Henry G. Wilcox Malay Gandhi Micheal H. Bahr Micheal C. Allan Linus A. Wodi George A. Cook Thomas A. Hughes Rajendra Raghow 《Obesity (Silver Spring, Md.)》2009,17(8):1563-1573
The objective of this study was to determine the molecular bases of disordered hepatic function and disease susceptibility in obesity. We compared global gene expression in liver biopsies from morbidly obese (MO) women undergoing gastric bypass (GBP) surgery with that of women undergoing ventral hernia repair who had experienced massive weight loss (MWL) following prior GBP. Metabolic and hormonal profiles were examined in MO vs. MWL groups. Additionally, we analyzed individual profiles of hepatic gene expression in liver biopsy specimens obtained from MO and MWL subjects. All patients underwent preoperative metabolic profiling. RNAs were extracted from wedge biopsies of livers from MO and MWL subjects, and analysis of mRNA expression was carried out using Affymetrix HG‐U133A microarray gene chips. Genes exhibiting greater than twofold differential expression between MO and MWL subjects were organized according to gene ontology and hierarchical clustering, and expression of key genes exhibiting differential regulation was quantified by real‐time–polymerase chain reaction (RT‐PCR). We discovered 154 genes to be differentially expressed in livers of MWL and MO subjects. A total of 28 candidate disease susceptibility genes were identified that encoded proteins regulating lipid and energy homeostasis (PLIN, ENO3, ELOVL2, APOF, LEPR, IGFBP1, DDIT4), signal transduction (MAP2K6, SOCS‐2), postinflammatory tissue repair (HLA‐DQB1, SPP1, P4HA1, LUM), bile acid transport (SULT2A, ABCB11), and metabolism of xenobiotics (GSTT2, CYP1A1). Using gene expression profiling, we have identified novel candidate disease susceptibility genes whose expression is altered in livers of MO subjects. The significance of altered expression of these genes to obesity‐related disease is discussed. 相似文献
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