Abstract：
:Crystallographic analysis revealed that the nicotinamide ring of NAD can bind with multiconformations to aldehyde dehydrogenase (ALDH) (Ni, L., Zhou, J., Hurley, T. D., and Weiner, H. (1999) Protein Sci. 8, 2784-2790). Electron densities can be defined for two conformations, neither of which appears to be compatible with the catalytic reaction. In one conformation, it would prevent glutamate 268 from functioning as a general base needed to activate the catalytic nucleophile, cysteine 302. In the other conformation, the nicotinamide is too far from the enzyme-substrate adduct for efficient hydride transfer. In this study, NMR and fluorescence spectroscopies were used to demonstrate that NAD and NADH bind to human liver cytosol and mitochondrial ALDH such that the nicotinamide samples a population of conformations while the adenosine region remains relatively immobile. Although the nicotinamide possesses extensive conformational heterogeneity, the catalyzed reaction leads to the stereospecific transfer of hydride to the coenzyme. Mobility allows the nicotinamide to move into position to be reduced by the enzyme-substrate adduct. Although the reduced nicotinamide ring retains mobility after NADH formation, the extent of the motion is less than that of NAD. It appears that after reduction the population of favored nicotinamide conformations shifts toward those that do not interfere with the ability of the enzyme to release the reaction product. In the case of the mitochondrial, but not the cytosolic, enzyme this change in conformational preference is promoted by the presence of Mg2+ ions. Coenzyme conformational mobility appears to be beneficial to catalysis by ALDH throughout the catalytic cycle.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Hammen PK,Allali-Hassani A,Hallenga K,Hurley TD,Weiner Hdoi
10.1021/bi012197tsubject
Has Abstractpub_date
2002-06-04 00:00:00pages
7156-68issue
22eissn
0006-2960issn
1520-4995pii
bi012197tjournal_volume
41pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract：:A heparin-induced conformational change is required to convert antithrombin from a slow to a fast inhibitor of factor Xa. It has been proposed [van Boeckel et al. (1994) Nat. Struct. Biol. 1, 423-425] that the reactive center residue P14 is inserted into beta-sheet A in native antithrombin and is displaced from the be...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9604643
更新日期：1996-07-02 00:00:00
abstract：:Photosensitized oxidation reactions damage tissue by catalyzing the formation of oxyradicals and singlet oxygen. beta-Carotene is hypothesized to exert photoprotective effects by quenching singlet oxygen formed by Type II reactions and by scavenging free radicals formed by Type I reactions. beta-Carotene antioxidant m...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9708646
更新日期：1997-10-21 00:00:00
abstract：:A method is described for measuring equilibrium constants of DNA-protein interactions using gel chromatography. This technique has been used to study the sequence-specific interaction of the HinfI restriction endonuclease with DNA. HinfI has a monomeric molecular weight of 31000 and exists as a dimer in its active for...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00333a037
更新日期：1985-06-04 00:00:00
abstract：:A covalent dimer of Saccharomyces cerevisiae iso-1 cytochrome c is stabilized by an interchain disulfide bond involving the cysteine residue penultimate to the C-terminus. The individual chains in the dimer appear to retain the tertiary structural features characteristic for monomeric cytochrome c albeit with some per...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00335a011
更新日期：1985-07-02 00:00:00
abstract：:The organization and interaction of chlorophylls (Chl) and the kinetics of the energy transfer in the core antenna of photosystem I (PSI) trimeric and monomeric complexes, isolated from Spirulina platensis with Triton X-100 have been studied by stationary and time-resolved fluorescence. At 295 K both complexes show an...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi970386z
更新日期：1997-11-11 00:00:00
abstract：:Cadherins are single pass transmembrane glycoproteins which mediate calcium dependent cell-cell adhesion by homophilic interactions. To reveal the molecular details of calcium binding and homoassociation, we recombinantly expressed in Escherichia coli a domain pair consisting of the first two domains of E-cadherin (EC...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9705624
更新日期：1997-06-24 00:00:00
abstract：:The gene for the human substance P receptor (NK-1) was cloned using cDNA probes made by the polymerase chain reaction from primers based on the rat sequence. The gene spans 45-60 kb and is contained in five exons, with introns interrupting at sites homologous to those in the NK-2 receptor gene. Analysis of restriction...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00108a006
更新日期：1991-11-05 00:00:00
abstract：:The qualitative and quantitative features of mutagenesis by two DNA adducts of carcinogenic alkylating agents, O6-methylguanine (m6G) and O4-methylthymine (m4T), were examined in vivo. The deoxyhexanucleotides 5'-GCTAGC-3' and 5'-GCTAGC-3' were synthesized, where the underlined bases are the positions of m4T or m6G, r...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00242a031
更新日期：1991-07-16 00:00:00
abstract：:Thermodynamic and structural evidence indicates that the DNA binding domains of lac repressor (lacI) exhibit significant conformational adaptability in operator binding, and that the marginally stable helix-turn-helix (HTH) recognition element is greatly stabilized by operator binding. Here we use circular dichroism a...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0270992
更新日期：2003-02-25 00:00:00
abstract：:Complex I of respiratory chains transfers electrons from NADH to ubiquinone, coupled to the translocation of protons across the membrane. Two alternative coupling mechanisms are being discussed, redox-driven or conformation-driven. Using "zero-length" cross-linking reagent and isolated hydrophilic domains of complex I...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi801160u
更新日期：2008-09-30 00:00:00
abstract：:Neuraminidase is one of the two glycoprotein spikes protruding from the influenza virus membrane. We have determined by X-ray crystallography the native structure of B/Lee/40 neuraminidase (NA) and the structures of its crystals soaked with a substrate, N-acetylneuraminyllactose (NANL), and an inhibitor, 2-deoxy-2,3-d...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00193a002
更新日期：1994-07-12 00:00:00
abstract：:Secondary structure in RNA targets is a significant barrier to short DNA probes. However, when such targets are the end product of an in vitro amplification scheme, it is possible to carry out transcription in the presence of nucleoside triphosphate analogues that reduce secondary structure of the RNA without impairin...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi049196w
更新日期：2004-08-10 00:00:00
abstract：:Binding of mitochondrial cyclophilin (CyP) to the inner mitochondrial membrane is induced by treatment of mitochondria with thiol reagents or oxidative stress and correlates with a sensitization to [Ca2+] of the cyclosporin A-sensitive mitochondrial permeability transition pore (MTP) [Connern, C. P., & Halestrap, A. P...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9525177
更新日期：1996-06-25 00:00:00
abstract：:The Synechococcus PCC7942 SmtB is a zinc-responsive transcriptional repressor and a member of the ArsR superfamily of prokaryotic metalloregulatory transcription factors. The mechanism of negative regulation by Zn(II) and other metals as well as the coordination chemistry (stoichiometry, affinity, and specificity) of ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi001140o
更新日期：2000-09-26 00:00:00
abstract：:Seipin is a transmembrane protein that resides in the endoplasmic reticulum and concentrates at junctions between the ER and cytosolic lipid droplets. Mutations in the human seipin gene, including the missense mutation A212P, lead to congenital generalized lipodystrophy (CGL), characterized by the lack of normal adipo...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi1013003
更新日期：2010-12-21 00:00:00
abstract：:A wide range of two-dimensional 1H NMR experiments have been used to completely assign the 500-MHz 1H NMR spectrum of recombinant Ca2+-saturated bovine calbindin D9k (76 amino acids, Mr = 8500). In solution, calbindin D9k exists as an equilibrium mixture of isoforms with trans (75%) and cis (25%) isomers of the peptid...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00443a043
更新日期：1989-08-22 00:00:00
abstract：:The time-resolved fluorescence decay of plant PSI-LHCI has been analyzed and compared with its component parts, the PSI core and the peripheral antenna LHCI, in an attempt to (i) define the physical domains associated with the multicomponent decay-associated spectra (DAS) and determine the origin of the kinetically sl...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi060243p
更新日期：2006-06-06 00:00:00
abstract：:Segregation of mRNAs in the cytoplasm of polar cells has been demonstrated for proteins involved in Xenopus and Drosophila oogenesis, and for some proteins in somatic cells. It is assumed that vectorial transport of the messages is generally responsible for this localization. The mRNA encoding the basic protein of cen...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9800247
更新日期：1998-05-12 00:00:00
abstract：:Regulation of cellular response is an important mechanism for controlling cellular functions. The transmembrane signaling of the hormone receptors is regulated by GTP-binding proteins (GTPases) and their associated proteins. Our previous studies demonstrated that the bifunctional GTP-binding protein, G alpha h (transg...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9522965
更新日期：1996-02-27 00:00:00
abstract：:We describe the synthesis of bordifluoropyrromethene (BODIPY), fluorescein, and related fluorescent derivatives of the beta-adrenergic ligand CGP 12177. With these probes we screened insect (Sf9) cells stably transformed with the human beta 2-adrenoceptor gene and expressing (2-3.5) x 10(5) human beta 2-adrenoceptors ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00197a015
更新日期：1994-08-09 00:00:00
abstract：:The ability of the low molecular weight lung surfactant-associated proteins, SP-B and SP-C, to alter the thermotropic properties of synthetic multilamellar vesicles was tested using differential scanning calorimetry (DSC). The presence of either SP-B or SP-C in dipalmitoylphosphatidylcholine (DPPC) or dipalmitoylphosp...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00053a026
更新日期：1993-01-19 00:00:00
abstract：:Determining the sequence of bond cleavages, and consequently the nature of intermediates, in enzyme-catalyzed reactions is a major goal of mechanistic enzymology. When significant primary isotope effects on V/K are observed for two different bond cleavages, both bonds may be broken in the same transition state or they...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00238a013
更新日期：1991-06-18 00:00:00
abstract：:Vitronectin is a multifunctional plasma glycoprotein which may regulate the systems related to protease cascades such as the coagulation, fibrinolysis, and complement systems as well as cell adhesion. Solid-phase assays and affinity chromatography on immobilized glycolipids indicated that vitronectin purified under de...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi972247n
更新日期：1998-05-05 00:00:00
abstract：:Deamidation of proteins is one of the most prevalent post-translational modifications found upon aging, and in age-onset diseases. Specific asparagine and glutamine residues are often selectively deamidated during this process. In the human lens, deamidation has been shown to occur in many crystallins, but it is not c...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.5b00185
更新日期：2015-08-11 00:00:00
abstract：:Lactate dehydrogenase (LDH) catalyzes the interconversion between pyruvate and lactate with nicotinamide adenine dinucleotide (NAD) as a cofactor. Using isotope-edited difference Fourier transform infrared spectroscopy on the "live" reaction mixture (LDH·NADH·pyruvate ⇌ LDH·NAD(+)·lactate) for the wild-type protein an...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi500215a
更新日期：2014-03-25 00:00:00
abstract：:The gamma-phosphoryl groups of two intermediates (M-ATP and M-ADP-P1) in the pathway of MgATP hydrolysis by myosin undergo extensive oxygen exchange with water. Actin activates the overall rate of hydrolysis at a rate-limiting step which follows these exchange reactions. Thus, actin, by decreasing the turnover time of...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00620a022
更新日期：1977-01-11 00:00:00
abstract：:1H NMR studies on the nonselective endothelin receptor agonist sarafotoxin SRTb have identified a helix between residues Asp 8 and His 16, and a beta-turn involving residues Cys 3 to Met 6; however, the biologically important C-terminal five residues were found to be conformationally variable. The average RMSD, measur...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00006a024
更新日期：1995-02-14 00:00:00
abstract：:Synaptotagmin 1 (syt1) is a synaptic vesicle-anchored membrane protein that acts as the calcium sensor for the synchronous component of neuronal exocytosis. Using site-directed spin labeling, the position and membrane interactions of a fragment of syt1 containing its two C2 domains (syt1C2AB) were assessed in bilayers...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi200049c
更新日期：2011-04-05 00:00:00
abstract：:Spectroscopic and kinetic methods have been used to explore the roles of divalent metal ions in the enolase-catalyzed dehydration of 2-phosphoglycerate (2-PGA). Enolase requires 2 equiv of metal ion per active site for maximal activity. Previous crystallographic studies [Larsen, T. M., Wedekind, J. E., Rayment, I., an...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0103922
更新日期：2001-07-10 00:00:00
abstract：:The DeltaG degrees (N)(-)(D) value obtained from extrapolation to zero denaturant concentration by the linear extrapolation method (LEM) is commonly interpreted to represent the Gibbs energy difference between native (N) and denatured (D) ensembles at the limit of zero denaturant concentration. For DeltaG degrees (N)(...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi001071d
更新日期：2000-12-12 00:00:00
