Abstract：
:Large primary deuterium kinetic isotope effects (1° DKIEs) on enzyme-catalyzed hydride transfer may be observed when the transferred hydride tunnels through the energy barrier. The following 1° DKIEs on kcat/ Km and relative reaction driving force are reported for wild-type and mutant glycerol-3-phosphate dehydrogenase (GPDH)-catalyzed reactions of NADL (L = H, D): wild-type GPDH, ΔΔ G ⧧ = 0 kcal/mol, 1° DKIE = 1.5; N270A, 5.6 kcal/mol, 3.1; R269A, 9.1 kcal/mol, 2.8; R269A + 1.0 M guanidine, 2.4 kcal/mol, 2.7; R269A/N270A, 11.5 kcal/mol, 2.4. Similar 1° DKIEs were observed on kcat. The narrow range of 1° DKIEs (2.4-3.1) observed for a 9.1 kcal/mol change in reaction driving force provides strong evidence that these are intrinsic 1° DKIEs on rate-determining hydride transfer. Evidence is presented that the intrinsic DKIE on wild-type GPDH-catalyzed reduction of DHAP lies in this range. A similar range of 1° DKIEs (2.4-2.9) on ( kcat/ KGA, M-1 s-1) was reported for dianion-activated hydride transfer from NADL to glycolaldehyde (GA) [Reyes, A. C.; Amyes, T. L.; Richard, J. P. J. Am. Chem. Soc. 2016, 138, 14526-14529]. These 1° DKIEs are much smaller than those observed for enzyme-catalyzed hydrogen transfer that occurs mainly by quantum mechanical tunneling. These results support the conclusion that the rate acceleration for GPDH-catalyzed reactions is due to the stabilization of the transition state for hydride transfer by interactions with the protein catalyst. The small 1° DKIEs reported for mutant GPDH-catalyzed and for wild-type dianion-activated reactions are inconsistent with a model where the dianion binding energy is utilized in the stabilization of a tunneling ready state.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Reyes AC,Amyes TL,Richard JPdoi
10.1021/acs.biochem.8b00536subject
Has Abstractpub_date
2018-07-24 00:00:00pages
4338-4348issue
29eissn
0006-2960issn
1520-4995journal_volume
57pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract：:Ferredoxin I from Desulfovibrio africanus (Da FdI) is a small acidic [4Fe-4S] cluster protein that exchanges electrons with pyruvate-ferredoxin oxidoreductase (PFOR), a key enzyme in the energy metabolism of anaerobes. The thermodynamic properties and the electron transfer between PFOR and either native or mutated FdI...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0485878
更新日期：2004-12-14 00:00:00
abstract：:The complete amino acid sequence was determined for bovine ubiquitin, and adenylate cyclase stimulating polypeptide, which is probably represented universally in living cells. Ubiquitin has a molecular weight of 8451 and consists of a single polypeptide chain containing 74 amino acid residues. It contains four argini...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00681a026
更新日期：1975-05-20 00:00:00
abstract：:Members of the uracil-DNA glycosylase (UDG) enzyme family recognize and bind uracil, sequestering it within the binding site pocket and catalyzing the cleavage of the base from the deoxyribose, leaving an abasic site. The recognition and binding are passive and rely on innate dynamic motions of DNA wherein base pairs ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.8b00676
更新日期：2018-10-02 00:00:00
abstract：:The beta-glycosidase from the hyperthermophilic Archaeon Sulfolobus solfataricus hydrolyzes beta-glycosides following a retaining mechanism based upon the action of two amino acids: Glu387, which acts as the nucleophile of the reaction, and Glu206, which acts as the general acid/base catalyst. The activities of inacti...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi981855f
更新日期：1998-12-08 00:00:00
abstract：:Taurine/alpha-ketoglutarate (alphaKG) dioxygenase (TauD), an archetype alphaKG-dependent hydroxylase, is a non-heme mononuclear Fe(II) enzyme that couples the oxidative decarboxylation of alphaKG with the conversion of taurine to aminoacetaldehyde and sulfite. The crystal structure of taurine-alphaKG-Fe(II)TauD is kno...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi048746n
更新日期：2005-03-15 00:00:00
abstract：:Folylpolyglutamate synthetase was purified 30,000-150,000-fold from hog liver. Purification required the use of protease inhibitors, and the protein was purified to homogeneity in two forms. Both forms of the enzyme were monomers of Mr 62,000 and had similar specific activities. The specific activity of the homogeneou...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00376a024
更新日期：1987-01-27 00:00:00
abstract：:We measure the effects of low concentrations of helix-stabilizing cosolvents, including 2,2,2-trifluoroethanol (TFE), on the thermodynamics and kinetics of folding of the dimeric alpha-helical coiled coil derived from the leucine zipper region of bZIP transcriptional activator GCN4. The change in kinetic behavior upon...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi981641y
更新日期：1998-10-13 00:00:00
abstract：:Designing subtype-selective agonists for neuronal nicotinic acetylcholine receptors is a challenging and significant goal aided by intricate knowledge of each subtype's binding patterns. We previously reported that in α6β2 receptors, acetylcholine makes a functional cation-π interaction with Trp149, but nicotine and T...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.7b00213
更新日期：2017-04-04 00:00:00
abstract：:Spectrin dimers interact weakly with F-actin under physiological solvent conditions (with an association constant of about 5 X 10(3) M-1 at 20 degrees C). In the presence of the membrane skeletal constituent, protein 4.1, strong binding is observed; an analysis of the profiles for formation of a ternary complex leads ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00314a027
更新日期：1984-09-11 00:00:00
abstract：:Cyanobacteriochromes (CBCRs) are cyanobacterial photoreceptors distantly related to phytochromes. Like phytochromes, CBCRs photointerconvert between two photostates that accompany photoisomerization of their bilin chromophores. While phytochromes typically exhibit red/far-red photocycles, CBCR photocycles are much mor...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi4015538
更新日期：2014-02-18 00:00:00
abstract：:The activation of adenosine-5'-monophosphate aminohydrolase from rabbit skeletal muscle by H+ has been demonstrated. Evidence is presented which indicates that the binding of H+ and K+ is linked, in that the dissociation constant (KA) for K+ activation is reduced as the pH is lowered. Concomitantly, the pK of several ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00641a013
更新日期：1977-11-01 00:00:00
abstract：:11Beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) catalyzes the conversion of 11-dehydrocorticosterone to its active form corticosterone in rodents (or cortisone to cortisol in humans). The reductive reaction of the 11-keto to 11-hydroxyl is the pivotal switch in the activation of glucocorticoids. An excess of ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi047599q
更新日期：2005-05-10 00:00:00
abstract：:Over 155 mutations within the V2 vasopressin receptor (AVPR2) gene are responsible for nephrogenic diabetes insipidus (NDI). The expression and subcellular distribution of four of these was investigated in transfected cells. These include a point mutation in the seventh transmembrane domain (S315R), a frameshift mutat...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi002699r
更新日期：2001-06-12 00:00:00
abstract：:The present study was carried out to determine the energetics of Clostridium symbiosum pyruvate phosphate dikinase (PPDK) catalyzed interconversion of adenosine 5'-triphosphate (ATP), orthophosphate (Pi), and pyruvate (pyr) with adenosine 5'-monophosphate (AMP), inorganic pyrophosphate, and phosphoenolpyruvate (PEP) [...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00171a007
更新日期：1994-02-08 00:00:00
abstract：:Bioluminescence with luciferase-luciferin pairs is an attractive method for surveying cells in live tissues and whole organisms. Recent advances in luciferin chemistry and luciferase engineering are further expanding the scope of the technology. It is now possible to spy on cells in a variety of deep tissues and visua...
journal_title：Biochemistry
pub_type： 杂志文章,评审
doi：10.1021/acs.biochem.7b00435
更新日期：2017-10-03 00:00:00
abstract：:ROS-GC represents a membrane guanylate cyclase subfamily whose distinctive feature is that it transduces diverse intracellularly generated Ca(2+) signals into the production of the second messenger cyclic GMP. An intriguing feature of the first subfamily member, ROS-GC1, is that it is both stimulated and inhibited by ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi990851n
更新日期：1999-09-28 00:00:00
abstract：:There are a small number of peptides derived from proteins that have a propensity to adopt structure in aqueous solution which is similar to the structure they possess in the parent protein. There are far fewer examples of protein fragments which adopt stable nonnative structures in isolation. Understanding how nonnat...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi990320z
更新日期：1999-06-01 00:00:00
abstract：:In halophilic archaea the photophobic response is mediated by the membrane-embedded 2:2 photoreceptor/-transducer complex SRII/HtrII, the latter being homologous to the bacterial chemoreceptors. Both systems bias the rotation direction of the flagellar motor via a two-component system coupled to an extended cytoplasmi...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi501160q
更新日期：2015-01-20 00:00:00
abstract：:Oligonucleotides capable of sequence-specific triple helix formation have been proposed as DNA binding ligands useful for modulation of gene expression and for directed genome modification. However, the effectiveness of such triplex-forming oligonucleotides (TFOs) depends on their ability to bind to their target sites...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi982290q
更新日期：1999-02-09 00:00:00
abstract：:The N-terminal region of alpha-spectrin is responsible for its association with beta-spectrin in a heterodimer, forming functional tetramers. Non-erythroid alpha-spectrin (alphaII-spectrin) has a significantly higher association affinity for beta-spectrin than the homologous erythroid alpha-spectrin (alphaI-spectrin)....
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi8013032
更新日期：2009-01-13 00:00:00
abstract：:The first indication of the size of a conformational change implicated in ion transport by sodium pump has been obtained by measuring the change in efficiency of fluorescence energy transfer between two specific locations on the alpha-subunit. The donor (5'-(iodoacetamido)fluorescein) attaches covalently to cysteine-4...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi960407+
更新日期：1996-06-25 00:00:00
abstract：:The calcium binding protein alpha-lactalbumin folds via a molten globule intermediate. Calcium does not bind strongly to the unfolded protein or the molten globule, but does bind to the transition state between the molten globule and the native protein. Of interest are the structures formed in the transition state tha...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi962901j
更新日期：1997-04-15 00:00:00
abstract：:The membrane-proximal external region (MPER) of the gp41 fusion protein of HIV is highly conserved among isolates of this virus and is considered a target for vaccine development. This region also appears to play a role in membrane fusion as well as localization of the virus to cholesterol-rich domains in membranes. T...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi8014828
更新日期：2008-11-11 00:00:00
abstract：:Living cells contain a range of densely phosphorylated surfaces, including phospholipid membranes, ribonucleoproteins, and nucleic acid polymers. Hyperphosphorylated surfaces also accumulate in neurodegenerative diseases as neurofibrillar tangles. We have synthesized and structurally characterized a precisely patterne...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi500599a
更新日期：2014-07-08 00:00:00
abstract：:The beta- and delta-subunits of the nicotinic acetylcholine receptor from Torpedo californica were covalently photolabeled at the synaptic surface with the ATP photoaffinity analogue [alpha-32P]-8-azido-ATP. The specificity of labeling for nucleotide binding sites was demonstrated by the saturation of labeling with in...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00080a002
更新日期：1993-07-27 00:00:00
abstract：:Site-directed mutagenesis and high-resolution two-dimensional (2D) proton nuclear magnetic resonance (NMR) were used to probe the structural and functional roles of a highly conserved residue, Asp-49, in the interfacial catalysis by bovine pancreatic phospholipase A2 (PLA2, overexpressed in Escherichia coli). Accordin...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00253a009
更新日期：1994-12-13 00:00:00
abstract：:The kinetics of reduction of free flavin semiquinones of the individual components of 1:1 covalent and electrostatic complexes of yeast ferric and ferryl cytochrome c peroxidase and ferric horse cytochrome c have been studied. Covalent cross-linking between the peroxidase and cytochrome c at low ionic strength results...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00406a029
更新日期：1988-03-22 00:00:00
abstract：:Beta-lactams exert their antibiotic action through their inhibition of bacterial DD-peptidases (penicillin-binding proteins). Bacteria, in general, carry several such enzymes localized on the outside of their cell membrane to catalyze the final step in cell wall (peptidoglycan) synthesis. They have been classified int...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi061804f
更新日期：2006-12-26 00:00:00
abstract：:Tritiated sodium borohydride was used to reduce the substrates of triosephosphate isomerase in the presence of the enzyme, and the mixture of the four possible products (D-[1(R)-3H]; D-[1(S)-3H]-; D-[2-3H]-, and L-[2-3H]glycerol 3-phosphate) was analyzed. While enzyme-bound dihydroxyacetone phosphate is reduced comple...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00692a020
更新日期：1975-10-21 00:00:00
abstract：:A new actinoleukin-like antitumor antibiotic, BBM-928A, has been shown to interact with isolated DNA molecules. BBM-928A contains two substituted quinolines linked by a cyclic decapeptide. Quenching effects of the covalently closed superhelical PM2 DNA on the BBM-928A fluorescence revealed a strong interaction with an...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00565a012
更新日期：1980-11-25 00:00:00
