Abstract:
:Deletion of Glu146, Thr147, and Trp148 from thrombin dramatically alters its interactions with substrates, ligands, and inhibitors, and the changes appear to result from nonlocal modification of thrombin's structure [Le Bonniec, B. F., Guinto, E. R., & Esmon, C. T. (1992) J. Biol. Chem. 267, 19341-19348]. In an attempt to localize conformational change in this thrombin mutant (des-ETW), its affinity for 43 site-directed mutants of hirudin have been compared with that of native thrombin. The inhibition constants (KI) of recombinant and natural hirudin with des-ETW were found to be approximately 2300-fold higher than those with thrombin. The reduced affinity was predominantly the result of an increase in the dissociation rate constant rather than a decrease in the association rate constant. For most of the hirudin mutants tested, the difference in binding energy between thrombin and des-ETW [delta delta Gbo(IIa-ETW)] was about 19.9 kJ mol-1 and comparable to that of hirudin; in particular, all mutations in the C-terminal region of the inhibitor did not affect delta delta Gbo(IIa-ETW). Thus, the conformation of thrombin's anion-binding exosite seems unaltered by the des-ETW mutation. In contrast, hirudin substitutions involving Val1', Val2', Thr4', Asp5', and Ser19', as well as the addition of an N-terminal glycine, resulted in values of delta delta Gbo(IIa-ETW) which were 2 to 10 kJ mol-1 lower than that for hirudin. Furthermore, a Leu15'-->Ala hirudin mutant behaved with des-ETW, as a partial competitive inhibitor rather than a tight-binding inhibitor, in contrast to all other hirudin variants tested.(ABSTRACT TRUNCATED AT 250 WORDS)
journal_name
Biochemistryjournal_title
Biochemistryauthors
Le Bonniec BF,Betz A,Guinto ER,Esmon CT,Stone SRdoi
10.1021/bi00179a023subject
Has Abstractpub_date
1994-04-05 00:00:00pages
3959-66issue
13eissn
0006-2960issn
1520-4995journal_volume
33pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::The composition of the molybdate-stabilized glucocorticoid receptor (GR) complex has been investigated with a monoclonal antibody against the steroid-binding Mr 94 000 (94K) GR protein. It was concluded that one antibody molecule binds one 94K GR molecule. This finding constituted the basis for calculating the number ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00344a041
更新日期:1985-11-05 00:00:00
abstract::The crystal structure of Pseudomonas putida cytochrome P-450cam in the substrate-free form has been refined at 2.20-A resolution and compared to the substrate-bound form of the enzyme. In the absence of the substrate camphor, the P-450cam heme iron atom is hexacoordinate with the sulfur atom of Cys-357 providing one a...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00366a049
更新日期:1986-09-09 00:00:00
abstract::Metal homeostasis and resistance in bacteria is maintained by a panel of metal-sensing transcriptional regulators that collectively control transition metal availability and mediate resistance to heavy metal xenobiotics, including As(III), Cd(II), Pb(II), and Hg(II). The ArsR family constitutes a superfamily of metal ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi100490u
更新日期:2010-08-10 00:00:00
abstract::The interfacial interactions of cholesterol with sphingomyelins (SMs) containing various homogeneous acyl chains have been investigated by Langmuir film balance approaches. Low in-plane elasticity among the packed lipids was identified as an important physical feature of the cholesterol-sphingomyelin liquid-ordered ph...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi002791n
更新日期:2001-05-22 00:00:00
abstract::Compartmentation and NMR visibility of mitochondrial adenine nucleotides were quantitated in isolated rat liver mitochondria respiring on succinate and glutamate in vitro at 8 and 25 degrees C. Intra- and extramitochondrial nucleotides were discriminated by adding the chelator trans-1,2-diaminocyclohexane-N,N,N',N'-te...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00436a030
更新日期:1989-05-16 00:00:00
abstract::We systematically and quantitatively investigated the structure and thermodynamics of G-quadruplexes of RNAs and corresponding DNAs of the same sequences under molecular crowding conditions that mimic the high osmotic stress induced by the numerous molecules inside of living cells. Structural analyses demonstrated tha...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi1002822
更新日期:2010-06-01 00:00:00
abstract::Lysine side chains induce conformational changes in plasminogen (Pg) that regulate the process of fibrinolysis or blood clot dissolution. A lysine side-chain mimic, epsilon amino caproic acid (EACA), enhances the activation of Pg by urinary-type and tissue-type Pg activators but inhibits Pg activation induced by strep...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi992028x
更新日期:2000-04-25 00:00:00
abstract::Mercury ions can inhibit Candida albicans phosphomannose isomerase (PMI) by two different processes at sub-micromolar concentrations. Kinetic studies show that mercury ions are in rapid equilibrium with the enzyme and cause a clear partial noncompetitive inhibition when mannose 6-phosphate is used as the substrate. Th...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00190a018
更新日期:1994-06-21 00:00:00
abstract::The ribosome-dependent stringent factor reaction was found to be nonspecific with regard to the number of phosphate groups linked to the 5' position of guanosine nucleotides. Both GMP and guanosine 5'-tetraphosphate could accept a pyrophosphoryl group from ATP although at a much lower rate than GDP or GTP. Guanosine 5...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00676a015
更新日期:1975-03-11 00:00:00
abstract::The central importance of chorismate enzymes in bacteria, fungi, parasites, and plants combined with their absence in mammals makes them attractive targets for antimicrobials and herbicides. Two of these enzymes, anthranilate synthase (AS) and aminodeoxychorismate synthase (ADCS), are structurally and mechanistically ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.5b00013
更新日期:2015-04-14 00:00:00
abstract::c-Jun and c-Fos belong to the bZIP class of transcriptional activator proteins, many of which have been implicated in the neoplastic transformation of cells. We are interested in engineering dominant-negative leucine zipper (LZ) peptides as a means of sequestering these proteins in vivo in order to suppress their tran...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi952817o
更新日期:1996-07-16 00:00:00
abstract::We have covalently attached an electron paramagnetic resonance (EPR) spin probe to Cys-670 of the motor domain of ncd (nonclaret disjunctional protein) in order to investigate conformational changes associated with the chemomechanical cycle. Spin-labeling is highly specific and does not affect ncd function as monitore...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9706881
更新日期:1997-08-12 00:00:00
abstract::12-Iodo-cis-9-octadecenoic acid (12-IODE) is a time-dependent, irreversible inactivator of soybean lipoxygenase 1. The rate of inactivation is independent of 12-IODE concentration above 20 microM and is half-maximal at about 4 microM. Inactivation by 12-IODE requires lipid hydroperoxide, which must be present even aft...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00424a019
更新日期:1988-11-29 00:00:00
abstract::A topoisomer gel retardation assay has been used to examine the topological requirements for the formation of open promoter complexes on DNA minicircles carrying sigma 54-dependent promoters. In the absence of intercalators, individual topoisomers carrying both the nifL and nifF promoters could be resolved as discrete...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9701179
更新日期:1997-10-07 00:00:00
abstract::We have employed high-performance liquid chromatography (HPLC) to investigate the syntheses of histones H1 and H1o as synchronized cells traverse from mitosis to S phase. Chinese hamster (line CHO) cells were synchronized by mitotic selection, and, at appropriate times, they were pulse labeled for 1 h with [3H]lysine....
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00329a031
更新日期:1985-04-09 00:00:00
abstract::Aromatic amino acid aminotransferase (ArAT) from Escherichia coli was overexpressed in E. coli cells, purified, and characterized. The enzyme was similar to aspartate aminotransferase (AspAT) of E. coli in many aspects, such as gross protein structure and spectroscopic properties. The reactions of pyridoxal 5'-phospha...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00096a036
更新日期:1993-11-16 00:00:00
abstract::The temperature dependence of biological rates at different scales (from individual enzymes to isolated organisms to ecosystem processes such as soil respiration and photosynthesis) is the subject of much historical and contemporary research. The precise relationship between the temperature dependence of enzyme rates ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.0c00530
更新日期:2020-09-29 00:00:00
abstract::Typical of many transcriptional regulatory proteins, the lambdoid bacteriophage repressors bind cooperatively to multiple sites on DNA. This cooperative binding is essential for establishment and maintenance of phage lysogeny. In the phage, two repressor homodimers, one bound at each of the adjacent operator sites, in...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi027318j
更新日期:2003-04-15 00:00:00
abstract::Methylmalonyl-CoA mutase from Propionibacterium shermanii is an adenosylcobalamin-dependent enzyme which catalyzes the reversible isomerization of methylmalonyl-CoA and succinyl-CoA. The rate of tritium loss from 5'-[3H]adenosylcobalamin during the enzymic reaction and the relative rates of tritium appearance in subst...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi961250o
更新日期:1996-09-10 00:00:00
abstract::The growth and shortening of microtubules in guanosine triphosphate-(GTP-) mediated dynamic instability has previously been observed to occur at rates which are remarkably variable (Gildersleeve et al., 1992, Chrétien et al., 1995). Neighboring microtubules observed simultaneously can grow or shorten at different rate...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi961070e
更新日期:1996-11-12 00:00:00
abstract::Big ET-1 and big ET-3 are precursor peptides which render endothelin-1 (ET-1) and endothelin-3 (ET-3) relatively unreactive and resistant to proteolytic cleavage. Big ET-1 is cleaved in vivo by ECE-1 (endothelin-converting enzyme), and big ET-3 is also cleaved but apparently to a significantly lesser extent by this en...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi981689b
更新日期:1999-02-09 00:00:00
abstract::Alpha-sarcin is an exquisitely specific ribonuclease that binds and cleaves a single phosphodiester bond in the large rRNA of the eukaryotic ribosome, inactivating it. To better understand this remarkable activity, the contributions of the active site residues (His 50, Glu 96, and His 137) to the conformational stabil...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi061273v
更新日期:2006-11-21 00:00:00
abstract::PT-ACRAMTU ([PtCl(en)(ACRAMTU-S)](NO(3))(2), en = ethane-1,2-diamine, ACRAMTU = 1-[2-(acridin-9-ylamino)ethyl]-1,3-dimethylthiourea) is a cytotoxic platinum-acridine conjugate previously shown to form adducts with the N3 endocyclic nitrogen of adenine in the DNA minor groove. This unusual observation and our prior det...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi050745n
更新日期:2005-08-23 00:00:00
abstract::Many DNA polymerases select their natural substrates, deoxy- as opposed to ribonucleoside triphosphates, with a selectivity greater than 10000-fold. The function of a highly conserved residue, Tyr416, in the palm domain of the parental enzyme, an exo(-) derivative of RB69 DNA polymerase (gp43), a member of the pol alp...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0202171
更新日期:2002-08-13 00:00:00
abstract::Kinetics studies in homogeneous aqueous solution showed that solubilized penicillin-binding protein 2a (sPBP2a) of methicillin-resistant Staphylococcus aureus (a bacterial DD-peptidase) was inhibited by the amphiphilic glycolipid antibiotic moenomycin. Inhibition at the peptidase site was determined by competition exp...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi982309p
更新日期:1999-08-10 00:00:00
abstract::The dynamic formation of stress granules (SGs), processing bodies (PBs), and related RNA organelles regulates diverse cellular processes, including the coordination of functionally connected messengers, the translational regulation at the synapse, and the control of viruses and retrotransposons. Recent studies have sh...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.8b00025
更新日期:2018-05-01 00:00:00
abstract::Xylose isomerase (E.C. 5.3.1.5) catalyzes the interconversion of aldose and ketose sugars and has an absolute requirement for two divalent cations at its active site to drive the hydride transfer rates of sugar isomerization. Evidence suggests some degree of metal movement at the second metal site, although how this m...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi049812o
更新日期:2004-06-01 00:00:00
abstract::The conformations of three peptide models of the one-disulfide and fully reduced forms of apamin were characterized by using nuclear magnetic resonance (NMR) spectroscopy. Apa-2 contains the native disulfide bond between Cys3 and Cys15 in apamin with the other two cysteines replaced by alanines. Apa-1 contains the nat...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00183a031
更新日期:1994-05-03 00:00:00
abstract::A series of diastereomeric dipeptide amides, containing an N-terminal L-lysyl residue and a C-terminal L- or D-amino acid with a derivatized aromatic ring on the side chain, was synthesized to determine the dependence of (1) the chirality of the N-terminal amino acid alpha-carbon and (2) the length of the N-terminal a...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00278a005
更新日期:1983-04-26 00:00:00
abstract::A cDNA designated hIIB1, representing the entire coding sequence of a P450 in the IIB gene family, was isolated from a human liver lambda gt11 library by using the rat IIB1 cDNA as a probe. The hIIB1 protein, deduced from the cDNA sequence, contained 491 amino acids, had a calculated molecular weight of 56,286, and di...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00444a029
更新日期:1989-09-05 00:00:00