Abstract:
:A new series of peptide inhibitors of human Factor VIIa (FVIIa) has been identified and affinity matured from naive and partially randomized peptide phage libraries selected against the immobilized tissue factor x Factor VIIa (TF x FVIIa) complex. These "A-series" peptides contain a single disulfide bond and a 13-residue minimal core required for maximal affinity. They are exemplified by peptide A-183 (EEWEVLCWTWETCER), which binds at a newly identified exosite on the FVIIa protease domain, described in the accompanying report [Roberge, M., Santell, L., Dennis, M. S., Eigenbrot, C., Dwyer, M. A., and Lazarus, R. A. (2001) Biochemistry 40, 9522-9531]. A-183 was obtained from a trypsin digest of A-100-Z, a recombinant protein comprising A-183 and the Z domain of protein A. Surprisingly, A-183 was a very potent inhibitor of TF x FVIIa, inhibiting activation of Factor X (FX) and Factor IX and amidolytic activity of Chromozym t-PA with IC50 values of 1.6 +/- 1.2, 3.5 +/- 0.3, and 8.5 +/- 3.5 nM, respectively. Kinetic analysis revealed that A-183 was a partial (hyperbolic) mixed-type inhibitor of FX activation having a Ki of 200 pM as well as a partial competitive inhibitor of amidolytic activity. The A-series peptides were also specific and potent inhibitors of TF-dependent clotting as measured in a prothrombin time (PT) clotting assay and had no effect on the TF-independent activated partial thromboplastin time. At saturating concentrations of peptide, the maximal extent by which A-183 and A-100-Z inhibited the rate of FX activation was 78 +/- 3 and 89 +/- 6%, respectively. The degree of inhibition of the rate of FX activation correlated with a maximum fold prolongation in the PT assay of 1.8-fold for A- 183 and 3.3-fold for A-100-Z. The A-series peptides represent a new class of peptide exosite inhibitors that are capable of attenuating, rather than completely inhibiting, the activity of TF x FVIIa, potentially leading to anticoagulants with an increased therapeutic window.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Dennis MS,Roberge M,Quan C,Lazarus RAdoi
10.1021/bi010591lsubject
Has Abstractpub_date
2001-08-14 00:00:00pages
9513-21issue
32eissn
0006-2960issn
1520-4995journal_volume
40pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::The objective of this study was to determine the molecular factors that lead to beta-lactamase inhibitor resistance for the M69V variant in SHV-1 beta-lactamase. With mechanism-based inhibitors, the beta-lactamase forms an acyl-enzyme intermediate that consists of a trans-enamine derivative in the active site. This st...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi060990m
更新日期:2006-10-03 00:00:00
abstract::Understanding the structural basis for defects in protein function that underlie protein-based genetic diseases is the fundamental requirement for development of therapies. This situation is epitomized by the cystic fibrosis transmembrane conductance regulator (CFTR)-the gene product known to be defective in CF patien...
journal_title:Biochemistry
pub_type: 杂志文章,评审
doi:10.1021/bi702209s
更新日期:2008-02-12 00:00:00
abstract::The correlation of the UVRR nuW3 mode with the tryptophan chi(2,1) dihedral angle [Maruyama and Takeuchi (1995) J. Raman Spectrosc. 26, 319; Miura et al. (1989) J. Raman Spectrosc. 20, 667; Takeuchi (2003) Biopolymers 72, 305] has been extended to a full, 360 degrees rotation. The 3-fold periodicity of the relationshi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi801293v
更新日期:2009-03-31 00:00:00
abstract::We have studied the Cu(II), Co(II), and Fe(III) complexes of the antineoplastic drug bleomycin by using electron spin--echo envelope spectroscopy. For all three complexes, nitrogen coordination of the metal ions is demonstrated. For the Cu(II)-- and Co(II)--drug complexes, we have been able to identify imidazole as a ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00509a045
更新日期:1981-03-17 00:00:00
abstract::In this work, we obtain the data needed to predict chemical interactions of polyethylene glycols (PEGs) and glycerol with proteins and related organic compounds and thereby interpret or predict chemical effects of PEGs on protein processes. To accomplish this, we determine interactions of glycerol and tetraEG with >30...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.5b00246
更新日期:2015-06-09 00:00:00
abstract::Recent studies of DNA hairpin loops have shown considerable dependence of the stability on the sequence of the loop [Senior, M., Jones, R. A., & Breslauer, K. J. (1988a) Proc. Natl. Acad. Sci. U.S.A. 85, 6242-6246; Xodo, L. E., Manzini, G., Quadrifoglio, F., van der Marel, G., & van Boom, J. H. (1989) Biochimie 71, 79...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00181a026
更新日期:1994-04-19 00:00:00
abstract::The interactions of Alzheimer's amyloid beta-peptide with cyclodextrins were studied by (1)H NMR: the translational diffusion coefficient of the peptide and chemical shift changes were studied by the presence of variable concentrations of cyclodextrins. For the full-length peptide, Abeta(1-40), the combined results of...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi036254p
更新日期:2004-05-25 00:00:00
abstract::The preferential in vitro methylation of histone H3 by coactivator-associated arginine methyltransferase 1 (CARM1) has been proposed as a basis for its ability to enhance gene transcription [Chen, D., et al. (1999) Science 284, 2174-2177]. To further evaluate the significance of H3 methylation, we studied the kinetics...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi002631b
更新日期:2001-05-15 00:00:00
abstract::Equilibrium binding experiments are widely used for the accurate characterization of binding and competitive binding behavior in biological systems. Modern high-throughput discovery efforts in chemical biology rely heavily upon this principle. Here, we derive exact analytical expressions for general competitive bindin...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi048233g
更新日期:2004-12-28 00:00:00
abstract::PDZ (acronym of the synapse-associated protein PSD-95/SAP90, the septate junction protein Discs-large, and the tight junction protein ZO-1) domains are abundant small globular protein interaction domains that mainly recognize the carboxyl termini of their target proteins. Detailed knowledge on PDZ domain binding speci...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi700954e
更新日期:2007-11-27 00:00:00
abstract::Biological membranes exhibit an asymmetric distribution of phospholipids. Phosphatidylserine (PS) is an acidic phospholipid that is found almost entirely on the interior of the cell where it is important for interaction with many cellular components. A less well understood phenomenon is the asymmetry of the neutral ph...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00119a022
更新日期:1992-02-04 00:00:00
abstract::Sialyltransferase activities, SAT-3 (CMP-NeuAc:nLcOse4Cer alpha 2-3sialyltransferase) and SAT-4 (CMP-NeuAc:GgOse4Cer alpha 2-3sialyltransferase), in Colo 205 cells catalyze the transfer of sialic acid to the terminal galactose of GlcNc-- and GalNAc-containing glycolipid substrates, respectively. Competition kinetic st...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi960239l
更新日期:1996-04-23 00:00:00
abstract::As a light-driven water-plastoquinone oxidoreductase, Photosystem II produces molecular oxygen as an enzymatic product. Additionally, under a variety of stress conditions, reactive oxygen species are produced at or near the active site for oxygen evolution. In this study, Fourier-transform ion cyclotron resonance mass...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi300650n
更新日期:2012-08-14 00:00:00
abstract::R67 dihydrofolate reductase (DHFR) is a plasmid-encoded, type II enzyme. Four monomers (78 amino acids long) assemble into a homotetramer possessing 222 symmetry. In previous studies, a tandem array of four R67 DHFR gene copies was fused in frame to generate a functional monomer named Quad1. This protein possessed the...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi1005943
更新日期:2010-08-31 00:00:00
abstract::Polyphosphate glucokinase from Mycobacterium tuberculosis catalyzes the phosphorylation of glucose using inorganic polyphosphates [poly(P)] or ATP. The steady-state kinetic mechanisms of the poly(P)- and ATP-dependent glucokinase reactions were investigated using initial velocity, product inhibition, and dead-end inhi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9528659
更新日期:1996-07-30 00:00:00
abstract::To identify the residues essential for interfacial binding and substrate binding of human pancreatic phospholipase A2 (hpPLA2), several ionic residues in the putative interfacial binding surface (R6E, K7E, K10E, and K116E) and substrate binding site (D53K and K56E) were mutated. Interfacial affinity of these mutants w...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi990600e
更新日期:1999-06-15 00:00:00
abstract::We have studied the competition between Na+ and K+ for coordination by G quartets using the oligonucleotide d(G3T4G3) as a model system. d(G3T4G3) forms a dimeric foldback structure containing three G quartets in the presence of either NaCl or KCl. Proton chemical shifts, which are particular to the species of coordin...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9620565
更新日期:1996-12-03 00:00:00
abstract::A faithful transcription system for ribosomal RNA genes has been developed by using components from the small free-living amoeba Acanthamoeba castellanii. The system utilizes protein-free recombinant DNA as a template and in addition requires a crude cell-free extract containing RNA polymerase I and a transcription in...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00313a025
更新日期:1984-08-28 00:00:00
abstract::Inhibition of gamma-aminobutyric acid aminotransferase (GABA-AT) increases the concentration of GABA, an inhibitory neurotransmitter in human brain, which could have therapeutic applications for a variety of neurological diseases, including epilepsy. On the basis of studies of several previously synthesized conformati...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi061592m
更新日期:2006-12-05 00:00:00
abstract::Barnase, the ribonuclease from Bacillus amyloliquefaciens, has been cloned and expressed in Escherichia coli [Hartley, R. W. (1988) J. Mol. Biol. 202, 913-915], thus enabling the overproduction and site-directed mutagenesis of one of the smallest enzymes (Mr equals 12,382). As barnase is also composed of just a single...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00435a033
更新日期:1989-05-02 00:00:00
abstract::A 31 amino acid fragment of the extracellular N-terminus of the human G-protein coupled receptor for parathyroid hormone (PTH1R) has been structurally characterized by NMR and molecular dynamics simulations. The fragment PTH1R[168-198] includes residues 173-189, shown by photoaffinity cross-linking to be a contact dom...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi981265h
更新日期:1998-09-15 00:00:00
abstract::Two major neutral glyceroglucolipids (A and B) have been isolated from lipid extract of human gastric content by the procedure involving column fractionation on DEAE-Sephadex, silicic acid, and thin-layer chromatography. Both glycolipids contained glucose, glyceryl ethers, and fatty acids. The structures of these glyc...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00637a003
更新日期:1977-09-06 00:00:00
abstract::Betaine-homocysteine S-methyltransferase (BHMT) is a zinc-dependent enzyme that catalyzes the transfer of a methyl group from glycine betaine (Bet) to homocysteine (Hcy) to form dimethylglycine (DMG) and methionine (Met). Previous studies in other laboratories have indicated that catalysis proceeds through the formati...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi049821x
更新日期:2004-05-11 00:00:00
abstract::Bacteriophage T7 codes for a single-stranded DNA binding protein. This protein is the product of gene 2.5 and has been found previously to stimulate specifically the activity of the phage-coded DNA polymerase. We report here that the T7 DNA binding protein also stimulates the activity of the phage-coded exonuclease. T...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00266a042
更新日期:1982-11-09 00:00:00
abstract::To investigate the effect of a reduced level of selenocysteine (Sec) tRNA[Ser]Sec in selenoprotein biosynthesis, two mouse embryonic stem (ES) cell lines heterozygous for the corresponding gene were generated by homologous recombination of the host genome with targeting vectors encoding a deleted or a disrupted tRNA[S...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi970608t
更新日期:1997-07-15 00:00:00
abstract::We have used a photochemical method to map covalent binding sites of the carcinogen benzo[a]pyrenediol epoxide (BPDE) within DNA from the transcriptional control region of the chicken adult beta-globin gene. Our preliminary low-resolution mapping has demonstrated that this region contains highly preferred BPDE binding...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00358a045
更新日期:1986-05-20 00:00:00
abstract::A panel of six naïve 14-residue random peptide libraries displayed polyvalently on M13 phage was pooled and sorted against human leukemia inhibitory factor (LIF). After four rounds of selection, a single large family of peptides with the consensus sequence XCXXXXG(A/S)(D/E)(W/F)WXCF was found to bind specifically to L...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi035303v
更新日期:2003-11-18 00:00:00
abstract::Phospholipid transfer protein (PLTP) is expressed by macrophage-derived foam cells in human atherosclerotic lesions, suggesting a regulatory role for PLTP in cellular cholesterol homeostasis. However, the exact role of PLTP in the reverse cholesterol transport pathway is not known. PLTP is present in plasma as two for...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi700833h
更新日期:2007-10-23 00:00:00
abstract::We have used resonance energy transfer to read out the interactions of the alpha subunit of transducin (alpha T) with the transducin beta gamma subunit complex (beta gamma T) and to compare the rate of aluminum fluoride-induced alpha T activation, as reflected by the enhancement of the alpha T tryptophan fluorescence,...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00199a046
更新日期:1994-08-23 00:00:00
abstract::Both activities of the aspartokinase--homoserine I (AK-HSD) of Escherichia coli are inhibited by threonine. Careful threonine binding studies have now been done which have allowed us to distinguish the various effects of threonine on the enzyme. The ultrafiltration technique of H. Paulus ((1969) Anal. Biochem. 32, 101...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00610a014
更新日期:1978-08-22 00:00:00