Abstract:
:Prothrombin is the precursor of thrombin, a central enzyme in coagulation. Autoantibodies to prothrombin are associated with thromboembolism, but the mechanisms by which the antibodies modulate the coagulation processes are not understood. We screened a panel of 34 monoclonal antibody light chains isolated from patients with multiple myeloma for prothrombinase activity by an electrophoresis method. Two light chains with the activity were identified, and one of the light chains was characterized further. The prothrombinase activity eluted from a gel-filtration column run in denaturing solvent (6 M guanidine hydrochloride) at the characteristic positions of the light chain dimer and monomer. A constant level of catalytic activity was observed across the width of the light chain monomer peak, assessed as the cleavage of IEGR-methylcoumarinamide, a peptide substrate corresponding to residues 268-271 of prothrombin. Hydrolysis of this peptide by the light chain was saturable and consistent with Michaelis-Menten-Henri kinetics (K(m) 103 microM; k(cat) of 2.62 x 10(-)(2)/min). Four cleavage sites in prothrombin were identified by N-terminal sequencing of the fragments: Arg(155)-Ser(156), Arg(271)-Thr(272), Arg(284)-Thr(285), and Arg(393)-Ser(394). The light chain did not cleave radiolabeled albumin, thyroglobulin, and annexin V under conditions that readily permitted detectable prothrombin cleavage. Two prothrombin fragments (M(r) 55 000 and 38 000), were isolated by anion-exchange chromatography and were observed to cleave a thrombin substrate, tosyl-GPR-nitroanilide. Conversion of fibrinogen to fibrin was accelerated by the prothrombin fragments generated by the light chain. These finding suggest a novel mechanism whereby antibodies can induce a procoagulant state, i.e., prothrombin activation via cleavage of the molecule.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Thiagarajan P,Dannenbring R,Matsuura K,Tramontano A,Gololobov G,Paul Sdoi
10.1021/bi992588wsubject
Has Abstractpub_date
2000-05-30 00:00:00pages
6459-65issue
21eissn
0006-2960issn
1520-4995pii
bi992588wjournal_volume
39pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::The kinetic and thermodynamic features of reactions catalyzed by present-day enzymes appear to be the consequence of the evolution of these proteins toward maximal catalytic effectiveness. These features are identified and analyzed (in detail for one substrate-one product enzymes) by using ideas that link the energeti...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00450a009
更新日期:1989-11-28 00:00:00
abstract::Thymidylate synthase from phage T4 (T4TS) is part of a complex of several enzymes required for coordinate DNA synthesis in infected Escherichia coli cells. It has been proposed that similar complexes of enzymes related to DNA synthesis are also functional in eukaryotes [Pardee, A. B. (1989) Science 246, 603-608]. To d...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00255a028
更新日期:1994-12-27 00:00:00
abstract::The dependence of the activity of bovine Cu,Zn superoxide dismutase on pH and ionic strength was extensively investigated in the ranges of pH 7.4-pH 12.3 and of ionic strength of 0.02-0.25 M. The results obtained indicate that two positively charged groups having pK values of approximately 10.1 and 10.8 are involved i...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00385a043
更新日期:1987-06-02 00:00:00
abstract::DNA-protein cross-links (DPCs) in nucleosome core particles (NCPs), the fundamental building block of chromatin, arise during times of cellular oxidative stress. These lesions are expected to be detrimental to the cell due to interference with processes like chromatin remodeling, transcription, DNA replication, and ep...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi700475b
更新日期:2007-09-25 00:00:00
abstract::Using solution peptide synthesis, we have made three series of N alpha-formylated homooligopeptides, from the dipeptide to the heptapeptide, derived from L-methionine, L-norleucine, and S-methyl-L-cysteine and related to the chemotactic peptide N alpha-formylmethionylleucylphenylalanine. Compounds were prepared to det...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00299a018
更新日期:1984-02-14 00:00:00
abstract::The interaction of proteoglycan monomers with hyaluronate in cartilage is mediated by a globular binding region at the N-terminus of the proteoglycan monomer; this interaction is stabilized by link protein. Sequences show that both the binding region (27% carbohydrate) and the link protein (6% carbohydrate) contain an...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00108a015
更新日期:1991-11-05 00:00:00
abstract::Structural differences in the [2Fe-2S] ferredoxin, putidaredoxin (Pdx), from the camphor hydroxylation pathway of Pseudomonas putida have been investigated as a function of oxidation state of the iron cluster. Pdx is involved in biological electron transfer to cytochrome P450(cam) (CYP101). Redox-dependent differences...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi050152c
更新日期:2005-06-28 00:00:00
abstract::Inosine 5'-monophosphate (IMP) cyclohydrolase catalyzes the cyclization of 5-formaminoimidazole-4-carboxamide ribonucleotide (FAICAR) to IMP in the final step of de novo purine biosynthesis. Two major types of this enzyme have been discovered to date: PurH in Bacteria and Eukarya and PurO in Archaea. The structure of ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi061637j
更新日期:2007-05-01 00:00:00
abstract::The ATP binding cassette (ABC) transporters ABCG2 and ABCB1 perform ATP hydrolysis-dependent efflux of structurally highly diverse compounds, collectively called allocrites. Whereas much is known about allocrite-ABCB1 interactions, the chemical nature and strength of ABCG2-allocrite interactions have not yet been asse...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.5b00649
更新日期:2015-10-13 00:00:00
abstract::A dark state tertiary structure in the cytoplasmic domain of rhodopsin is presumed to be the key to the restriction of binding of transducin and rhodopsin kinase to rhodopsin. Upon light-activation, this tertiary structure undergoes a conformational change to form a new structure, which is recognized by the above prot...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi010746p
更新日期:2001-10-23 00:00:00
abstract::Genomic clones containing the human Zn-alpha 2-glycoprotein gene were isolated. Two of them were for the functional gene overlapped, and the other four were for two different pseudogenes (1 and 2) retaining exon-intron organization. The complete DNA sequence of the functional gene (9.3 kb) and its 5'- and 3'-flanking ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00211a004
更新日期:1993-12-07 00:00:00
abstract::The intermediates in the oxidation of deoxyhemoglobin by ferricyanide in 0.1 M KCl, at 20 degrees C and three pH values, were studied by cryogenic techniques. Data analysis was carried out according to a simple four rate constant model, ignoring the functional heterogeneity of the subunits, to simulate the time course...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00070a035
更新日期:1993-05-18 00:00:00
abstract::Studies reveal that it is possible to design a palladium(II)-containing porphyrin to bind exclusively by intercalation to double-stranded DNA while simultaneously enhancing the ability to sensitize the formation of singlet oxygen. The comparisons revolve around the cations [5,10,15,20-tetra(N-methylpyridinium-4-yl)por...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi401610t
更新日期:2014-02-04 00:00:00
abstract::To characterize the role of water in protein-DNA interactions, we have studied the specificity of the EcoRI restriction endonuclease as a function of osmotic and hydrostatic pressure. The extent of cleavage by the enzyme at noncanonical ("star") sites is shown to depend uniquely upon the osmotic pressure in the reacti...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00179a001
更新日期:1994-04-05 00:00:00
abstract::Enzyme therapeutics that can degrade l-methionine (l-Met) are of great interest as numerous malignancies are exquisitely sensitive to l-Met depletion. To exhaust the pool of methionine in human serum, we previously engineered an l-Met-degrading enzyme based on the human cystathionine-γ-lyase scaffold (hCGL-NLV) to cir...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.6b01172
更新日期:2017-02-14 00:00:00
abstract::The preparation, spectroscopic investigation, structure determination, conformational analysis, and modeling of the Dewar pyrimidinone photoproduct of thymidylyl-(3'----5')-thymidine, previously referred to as TpT3 [Johns, H. E., Pearson, M. L., LeBlanc, J. C., & Heilleiner, C. W. (1964) J. Mol. Biol. 9, 503-524], is ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00419a007
更新日期:1988-09-20 00:00:00
abstract::An assay was developed to study the spontaneous transfer and transbilayer movement (flip-flop) of lipid analogs labeled with the fluorescent fatty acid, 5-(5,7-dimethyl BODIPY)-1-pentanoic acid (C5-DMB-) in large unilamellar lipid vesicles comprised of 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC). The assay is base...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi970145r
更新日期:1997-07-22 00:00:00
abstract::A new rapid procedure for purifying 10-formyltetrahydrofolate dehydrogenase results in 90 mg of pure enzyme from two rabbit livers. This abundant liver enzyme is known to bind its product tetrahydropteroylpentaglutamate (H4PteGlu5) so tightly that it does not dissociate during size exclusion chromatography. 10-Formylt...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9619684
更新日期:1996-12-10 00:00:00
abstract::Transport of hydrophilic di- and tripeptides into Lactococcus lactis is mediated by a proton motive force-driven peptide transport protein (DtpT) that shares similarity with eukaryotic peptide transporters, e.g., from kidney and small intestine of rabbit, man, and rat. Hydropathy profiling in combination with the "pos...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi963068t
更新日期:1997-06-03 00:00:00
abstract::Pseudomonas aeruginosa PA4872 was identified by sequence analysis as a structurally and functionally novel member of the PEP mutase/isocitrate lyase superfamily and therefore targeted for investigation. Substrate screens ruled out overlap with known catalytic functions of superfamily members. The crystal structure of ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi701954p
更新日期:2008-01-08 00:00:00
abstract::GGAs are a family of vesicle-coating regulatory proteins that function in intracellular protein transport. A GGA molecule contains four domains, each mediating interaction with other proteins in carrying out intracellular transport. The GAT domain of GGAs has been identified as the structural entity that binds membran...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi034334n
更新日期:2003-06-03 00:00:00
abstract::The proposal that microtubule dynamic instability results from stabilization of microtubule ends by tubulin-GDP-Pi subunits (where Pi is inorganic phosphate) [Melki et al. (1996) Biochemistry 35, 12038] was based on studies of GTP hydrolysis and microtubule assembly that showed that tubulin-GDP-Pi subunits can transie...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi972602+
更新日期:1998-09-15 00:00:00
abstract::Single-tryptophan-containing mutants of low adenylation state Escherichia coli glutamine synthetase (wild type has two tryptophans at positions 57 and 158) have been constructed and studied by multifrequency phase/modulation fluorescence spectroscopy. The W57L mutant (retains tryptophan at residue 158) and the W158S m...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00228a008
更新日期:1991-04-09 00:00:00
abstract::The complex of dioldehydrase with adenosylcobalamin (coenzyme B12) and potassium ion reacts with molecular oxygen in the absence of a substrate to oxidize coenzyme and inactivate the complex. In this article, high performance liquid chromatography and mass spectral analysis are used to identify the nucleoside products...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi700078z
更新日期:2007-06-19 00:00:00
abstract::Transport of protons and solutes across mitochondrial membranes is essential for many physiological processes. However, neither the proton-pumping respiratory chain complexes nor the mitochondrial secondary active solute transport proteins have been characterized electrophysiologically in their native environment. In ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi1011755
更新日期:2010-12-07 00:00:00
abstract::X-ray absorption spectra are reported for the multi-Cu oxidase Rhus vernicifera laccase in oxidized and fully reduced forms and for laccase from which the type 2 Cu has been depleted (T2D). The structure of the Cu K edge for both preparations shows the presence of CuII and CuI in the oxidized and reduced states, respe...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00310a008
更新日期:1984-07-17 00:00:00
abstract::The Bacillus subtilis DNA-dependent RNA polymerase holoenzyme and core enzyme each contain approximately two atoms of zinc per molecule. When the dissociated subunits of the enzyme are passed through a blue dextran-Sepharose affinity column, only the beta subunit binds to the column. The total zinc content of the enzy...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00632a012
更新日期:1977-06-28 00:00:00
abstract::Actin depolymerizing activity in serum can be attributed to the two proteins brevin and vitamin D binding protein (DBP). To investigate their mechanisms of action, we used a number of techniques, including procedures involving the fluorescent pyrene-labeled actin probe, to compare the interaction of the two proteins w...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00308a030
更新日期:1984-06-19 00:00:00
abstract::The nicotinic acetylcholine receptor (nAChR) from Torpedo marmorata carries two nonequivalent agonist binding sites at the alphadelta and alphagamma subunit interfaces. These sites have been characterized by time-resolved fluorescence with the partial nicotinic agonist dansyl-C(6)-choline (Dnscho). When bound to the d...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi992811p
更新日期:2000-06-13 00:00:00
abstract::Adriamycin plays a prominent role in the treatment of leukemia and solid tumors in man. The mode of interaction of adriamycin with its nuclear target, responsible for its therapeutic effect, is known [Berman, H. M., & Young, P.R. (1981) Annu. Rev. Biophys. Bioeng. 10, 87-114]. The planar anthracycline moiety of adriam...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00380a043
更新日期:1987-03-24 00:00:00