Purification, characterization, and fibrinogen cleavage sites of three fibrinolytic enzymes from the venom of Crotalus basiliscus basiliscus.

Abstract:

:Three distinct fibrinolytic enzymes have been purified from the venom of Crotalus basiliscus basiliscus (the Mexican west coast rattlesnake). The high-performance liquid chromatography-based purification comprised the following steps: (a) hydrophobic interaction chromatography; (b) hydroxylapatite chromatography; (c) anion-exchange chromatography. Following hydrophobic interaction chromatography two fibrinolytic activity peaks were detected, Cbfib1 and Cbfib2. Cbfib2 was rendered homogeneous following hydroxylapatite chromatography. Upon hydroxylapatite chromatography Cbfib1 was shown to consist of two components, Cbfib1.1 and Cbfib1.2. Both Cbfib1.1 and Cbfib1.2 were purified to homogeneity using anion-exchange chromatography. SDS-polyacrylamide gel electrophoresis revealed that Cbfib1.1 and Cbfib1.2 had similar molecular weights (approximately 23,500), whereas Cbfib2 displayed a molecular weight of approximately 22,500. The enzymes do not appear to be glycosylated. Tryptic digests of all three enzymes, analyzed by high-performance reverse-phase chromatography, suggest that Cbfib1.1 and Cbfib1.2 are closely related and different from Cbfib2. The latter displayed more similarity with Cbfib1.2 than with Cbfib1.1. Specific fibrinolytic activity for all three enzymes was very similar, but general proteolytic activity varied substantially with Cbfib2 showing a 12-fold higher specific proteolytic activity when compared to Cbfib1.1 and Cbfib1.2. None of these enzymes exhibited hemorrhagic activity when injected (up to 100 micrograms) subcutaneously into mice. Cbfib1.1 and Cbfib1.2 action against fibrinogen was directed equally against both the A alpha- and B beta-chains. Against fibrin the rate of degradation of the alpha-chain was considerably higher than that of the beta-chain. Cbfib2 showed mainly alpha-fibrin(ogen)ase activity with limited activity on the beta-chain. Several fibrinogen cleavage sites on the A alpha-chain have been identified: Cbfib1.1 and Cbfib1.2 cleave at Lys413-Leu414, Ser505-Thr506, and Tyr560-Ser561. Cbfib2 cleaves mainly at Gly254-Ser255 and Pro516-Met517.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Retzios AD,Markland FS Jr

doi

10.1021/bi00134a003

subject

Has Abstract

pub_date

1992-05-19 00:00:00

pages

4547-57

issue

19

eissn

0006-2960

issn

1520-4995

journal_volume

31

pub_type

杂志文章
  • X-ray crystallographic and biochemical characterization of the inhibitory action of an imidazole-dioxolane compound on heme oxygenase.

    abstract::Heme oxygenase (HO) catalyzes the regiospecific cleavage of the porphyrin ring of heme using reducing equivalents and O2 to produce biliverdin, iron, and CO. Because CO has a cytoprotective effect through the p38-MAPK pathway, HO is a potential therapeutic target in cancer. In fact, inhibition of the HO isoform HO-1 r...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi062264p

    authors: Sugishima M,Higashimoto Y,Oishi T,Takahashi H,Sakamoto H,Noguchi M,Fukuyama K

    更新日期:2007-02-20 00:00:00

  • The photosystem of Rhodobacter sphaeroides assembles with zinc bacteriochlorophyll in a bchD (magnesium chelatase) mutant.

    abstract::A Rhodobacter sphaeroides bchD (magnesium chelatase) mutant was studied to determine the properties of its photosystem in the absence of bacteriochlorophyll (BChl). Western blots of reaction center H, M, and L (RC H/M/L) proteins from mutant membranes showed levels of 12% RC H, 32% RC L, and 46% RC M relative to those...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi701407k

    authors: Jaschke PR,Beatty JT

    更新日期:2007-10-30 00:00:00

  • Structure and assembly of the heterotrimeric and homotrimeric C-propeptides of type I collagen: significance of the alpha2(I) chain.

    abstract::Assembly of the type I procollagen molecule begins with interactions among the C-pro alpha1(I) and C-pro alpha2(I) domains. The C-propeptide domains themselves have subdomains of distinct structures. The important questions are where chain association begins and the basis of the chain selectivity which leads to the pr...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi0508338

    authors: Malone JP,Alvares K,Veis A

    更新日期:2005-11-22 00:00:00

  • HlyC, the internal protein acyltransferase that activates hemolysin toxin: role of conserved histidine, serine, and cysteine residues in enzymatic activity as probed by chemical modification and site-directed mutagenesis.

    abstract::HlyC is an internal protein acyltransferase that activates hemolysin, a toxic protein produced by pathogenic Escherichia coli. Acyl-acyl carrier protein (ACP) is the essential acyl donor. Separately subcloned, expressed, and purified prohemolysin A (proHlyA), HlyC, and [1-14C]myristoyl-ACP have been used to study the ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi982491u

    authors: Trent MS,Worsham LM,Ernst-Fonberg ML

    更新日期:1999-03-16 00:00:00

  • Probing Structural Dynamics and Topology of the KCNE1 Membrane Protein in Lipid Bilayers via Site-Directed Spin Labeling and Electron Paramagnetic Resonance Spectroscopy.

    abstract::KCNE1 is a single transmembrane protein that modulates the function of voltage-gated potassium channels, including KCNQ1. Hereditary mutations in the genes encoding either protein can result in diseases such as congenital deafness, long QT syndrome, ventricular tachyarrhythmia, syncope, and sudden cardiac death. Despi...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.5b00505

    authors: Sahu ID,Craig AF,Dunagan MM,Troxel KR,Zhang R,Meiberg AG,Harmon CN,McCarrick RM,Kroncke BM,Sanders CR,Lorigan GA

    更新日期:2015-10-20 00:00:00

  • Activating anions that replace Cl- in the O2-evolving complex of photosystem II slow the kinetics of the terminal step in water oxidation and destabilize the S2 and S3 states.

    abstract::Photosystem II, the multisubunit protein complex that oxidizes water to O2, requires the inorganic cofactors Ca2+ and Cl- to exhibit optimal activity. Chloride can be replaced functionally by a small number of anionic cofactors (Br-, NO3-, NO2-, I-), but among these anions, only Br- is capable of restoring rates of ox...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi982295n

    authors: Wincencjusz H,Yocum CF,van Gorkom HJ

    更新日期:1999-03-23 00:00:00

  • Structure, spectroscopic, and redox properties of Rhodobacter sphaeroides reaction centers bearing point mutations near the primary electron donor.

    abstract::Single mutations of three amino acid residues in the vicinity of the primary electron donor, P, in the reaction center (RC) from Rhodobacter (Rb.) sphaeroides were constructed and characterized in order to study the effects of hydrogen-bonding on the physical properties of P. The mutations, Phe M197-->Tyr, Met L248-->...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00210a041

    authors: Wachtveitl J,Farchaus JW,Das R,Lutz M,Robert B,Mattioli TA

    更新日期:1993-11-30 00:00:00

  • Reactivity of horse liver alcohol dehydrogenase with 3-methylcyclohexanols.

    abstract::The specificity of horse liver alcohol dehydrogenase for cyclohexanol and its 3-methyl derivatives was investigated by stopped-flow and initial velocity kinetic studies. The (1S,3S)-3-methylcyclohexanol was 7 times more reactive (V/Km) than cyclohexanol, whereas the (1R,3R)-3-methylcyclohexanol was at least 1000 times...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00409a060

    authors: Lee KM,Dahlhauser KF,Plapp BV

    更新日期:1988-05-03 00:00:00

  • Structure of the ternary complex formed by a chemotaxis receptor signaling domain, the CheA histidine kinase, and the coupling protein CheW as determined by pulsed dipolar ESR spectroscopy.

    abstract::The signaling apparatus that controls bacterial chemotaxis is composed of a core complex containing chemoreceptors, the histidine autokinase CheA, and the coupling protein CheW. Site-specific spin labeling and pulsed dipolar ESR spectroscopy (PDS) have been applied to investigate the structure of a soluble ternary com...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi100055m

    authors: Bhatnagar J,Borbat PP,Pollard AM,Bilwes AM,Freed JH,Crane BR

    更新日期:2010-05-11 00:00:00

  • Secondary binding sites for triplex-forming oligonucleotides containing bulges, loops, and mismatches in the third strand.

    abstract::We have used DNase I footprinting to examine the binding of five different 17-mer oligonucleotides to a 53-base oligopurine tract containing four pyrimidine interruptions. Although all the expected triplexes formed with high affinity (K(d) approximately 10-50 nM), one oligonucleotide produced a footprint at a second s...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi992773+

    authors: Fox KR,Flashman E,Gowers D

    更新日期:2000-06-06 00:00:00

  • Selection of a carbohydrate-binding domain with a helix-loop-helix structure.

    abstract::We obtained a novel carbohydrate-binding peptide having a helix-loop-helix scaffold from a random peptide library. The helix-loop-helix peptide library randomized at five amino acid residues was displayed on the major coat protein of a filamentous phage. Affinity selection with a ganglioside, Galbeta1-3GalNAcbeta1-4(N...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi8000837

    authors: Matsubara T,Iida M,Tsumuraya T,Fujii I,Sato T

    更新日期:2008-07-01 00:00:00

  • Catalytic and DNA binding properties of the ogg1 protein of Saccharomyces cerevisiae: comparison between the wild type and the K241R and K241Q active-site mutant proteins.

    abstract::The Ogg1 protein of Saccharomyces cerevisiae belongs to a family of DNA glycosylases and apurinic/apyrimidinic site (AP) lyases, the signature of which is the alpha-helix-hairpin-alpha-helix-Gly/Pro-Asp (HhH-GPD) active site motif together with a conserved catalytic lysine residue, to which we refer as the HhH-GPD/K f...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi992262n

    authors: Guibourt N,Castaing B,Van Der Kemp PA,Boiteux S

    更新日期:2000-02-22 00:00:00

  • Calcium binding and homoassociation of E-cadherin domains.

    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

    authors: Koch AW,Pokutta S,Lustig A,Engel J

    更新日期:1997-06-24 00:00:00

  • Structural and functional properties of human hemoglobins reassembled after synthesis in Escherichia coli.

    abstract::Human hemoglobin produced in the Escherichia coli coexpression system of Hernan et al. [(1992) Biochemistry 31, 8619-8628] has been transformed into a functionally homogeneous protein whose properties closely approximate those of normal hemoglobin A. Both of the alpha and beta chains of this hemoglobin contain a valin...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi981986g

    authors: Hui HL,Kavanaugh JS,Doyle ML,Wierzba A,Rogers PH,Arnone A,Holt JM,Ackers GK,Noble RW

    更新日期:1999-01-19 00:00:00

  • Molecular basis of reduced albumin synthesis in Morris hepatoma 7777.

    abstract::The level of albumin mRNA in the normal Buffalo rat liver and Morris hepatoma 7777 was compared by molecular hybridization with albumin complementary DNA (cDNA) and translational assays. Albumin mRNA was found to be 10% of the total rat liver poly(A)-containing RNA population but reduced approximately fourfold in the ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00608a028

    authors: Tse TP,Morris HP,Taylor JM

    更新日期:1978-07-25 00:00:00

  • The structure of the bovine protein tyrosine phosphatase dimer reveals a potential self-regulation mechanism.

    abstract::The bovine protein tyrosine phosphatase (BPTP) is a member of the class of low-molecular weight protein tyrosine phosphatases (PTPases) found to be ubiquitous in mammalian cells. The catalytic site of BPTP contains a CX(5)R(S/T) phosphate-binding motif or P-loop (residues 12-19) which is the signature sequence for all...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi990381x

    authors: Tabernero L,Evans BN,Tishmack PA,Van Etten RL,Stauffacher CV

    更新日期:1999-09-07 00:00:00

  • Nuclear magnetic resonance studies of 2'- and 3'-ribonucleotide structures in solution.

    abstract::A systematic 220-MHz proton nuclear magnetic resonance (nmr) study has been made of all common purine and pyrimidine 2'(3')-ribonucleotides in D2O solutions at 20 plus or minus 2 degrees. Spectra for the entire series were measured under similar conditions of concentration, temperature, and ionic strength, thereby fac...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00674a013

    authors: Davies DB,Danyluk SS

    更新日期:1975-02-11 00:00:00

  • Photoreactions of the Histidine Kinase Rhodopsin Ot-HKR from the Marine Picoalga Ostreococcus tauri.

    abstract::The tiny picoalga, Ostreococcus tauri, originating from the Thau Lagoon is a member of the marine phytoplankton. Because of its highly reduced genome and small cell size, while retaining the fundamental requirements of a eukaryotic photosynthetic cell, it became a popular model organism for studying photosynthesis or ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.8b01200

    authors: Luck M,Velázquez Escobar F,Glass K,Sabotke MI,Hagedorn R,Corellou F,Siebert F,Hildebrandt P,Hegemann P

    更新日期:2019-04-09 00:00:00

  • Phase-resolved spectral measurements with several two tryptophan containing proteins.

    abstract::We have used frequency domain fluorescence techniques to resolve the component emission spectra for several two tryptophan containing proteins (e.g., horse liver alcohol dehydrogenase, sperm whale apomyoglobin, yeast 3-phosphoglycerate kinase, apoazurin from Alcaligenes denitricans). We have first performed multifrequ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00399a047

    authors: Eftink MR,Wasylewski Z,Ghiron CA

    更新日期:1987-12-15 00:00:00

  • A C-truncated glutamyl-tRNA synthetase specific for tRNA(Glu) is stimulated by its free complementary distal domain: mechanistic and evolutionary implications.

    abstract::Faithful translation of the genetic code is mainly based on the specificity of tRNA aminoacylation catalyzed by aminoacyl-tRNA synthetases. These enzymes are comprised of a catalytic core and several appended domains. Bacterial glutamyl-tRNA synthetases (GluRS) contain five structural domains, the two distal ones inte...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi801690f

    authors: Dubois DY,Blais SP,Huot JL,Lapointe J

    更新日期:2009-06-30 00:00:00

  • Identification of the serine residue phosphorylated by protein kinase C in vertebrate nonmuscle myosin heavy chains.

    abstract::Two-dimensional mapping of the tryptic phosphopeptides generated following in vitro protein kinase C phosphorylation of the myosin heavy chain isolated from human platelets and chicken intestinal epithelial cells shows a single radioactive peptide. These peptides were found to comigrate, suggesting that they were iden...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00218a012

    authors: Conti MA,Sellers JR,Adelstein RS,Elzinga M

    更新日期:1991-01-29 00:00:00

  • Properties of interacting aspartic acid and lysine residues in the lactose permease of Escherichia coli.

    abstract::The side chains of the interacting pair Asp237(helix VII)-Lys358(helix XI) or Asp240(helix VII)-Lys319(helix X) in the lactose permease of Escherichia coli were extended by replacement with Glu and/or Arg or by site-specific derivatization of single-Cys replacement mutants. Iodoacetic acid was used to carboxymethylate...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00089a019

    authors: Sahin-Tóth M,Kaback HR

    更新日期:1993-09-28 00:00:00

  • Conformational adaptation of apolipoprotein A-I to discretely sized phospholipid complexes.

    abstract::The conformational constraints for apoA-I bound to recombinant phospholipid complexes (rHDL) were attained from a combination of chemical cross-linking and mass spectrometry. Molecular distances were then used to refine models of lipid-bound apoA-I on both 80 and 96 A diameter rHDL particles. To obtain molecular const...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi700384t

    authors: Bhat S,Sorci-Thomas MG,Tuladhar R,Samuel MP,Thomas MJ

    更新日期:2007-07-03 00:00:00

  • Bimodal action of alkaline earth cations on Dictyostelium discoideum ribonuclease P activity.

    abstract::The ribonucleoprotein ribonuclease P (RNase P) cleaves all tRNA precursors endonucleolitically to produce the mature 5'-end. Dictyostelium discoideum RNase P displays an absolute requirement for Mg2+. Only the alkaline earth cations Ca2+, Sr2+, and Ba2+, under appropriate conditions can substitute to some extent for M...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi9808971

    authors: Tekos A,Stathopoulos C,Drainas D

    更新日期:1998-11-03 00:00:00

  • Glutamate 189 of the D1 polypeptide modulates the magnetic and redox properties of the manganese cluster and tyrosine Y(Z) in photosystem II.

    abstract::Recent models for water oxidation in photosystem II postulate that the tyrosine Y(Z) radical, Y(Z)(*), abstracts both an electron and a proton from the Mn cluster during one or more steps in the catalytic cycle. This coupling of proton- and electron-transfer events is postulated to provide the necessary driving force ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi992749w

    authors: Debus RJ,Campbell KA,Pham DP,Hays AM,Britt RD

    更新日期:2000-05-30 00:00:00

  • Conformational characteristics of luliberin. Luminescence properties at liquid-nitrogen temperature.

    abstract::The luminescence properties (fluorescence and phosphorescence) of luliberin have been investigated at liquid-nitrogen temperature (77 K) in 50% ethylene glycolaqueous buffer at various pH's. Calculation of the energy-transfer efficiency between Trp and Tyr residues leads to an evaluation of the distance separating the...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00671a009

    authors: Marche P,Montenay-Garestier T,Fromageot P,Hélène C

    更新日期:1976-12-28 00:00:00

  • An additional methyl group at the 10-position of retinal dramatically slows down the kinetics of the rhodopsin photocascade.

    abstract::The present study focuses on ligand-protein interactions in a rhodopsin analogue generated from bovine opsin and the 10-methyl homologue of 11-cis-retinal. The analogue pigment displays a reduced alpha-band at 506 +/- 2 and a stronger beta-band at 325 nm. Remarkably, the rotational strength of these bands observed in ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi972397y

    authors: DeLange F,Bovee-Geurts PH,VanOostrum J,Portier MD,Verdegem PJ,Lugtenburg J,DeGrip WJ

    更新日期:1998-02-03 00:00:00

  • Transmembrane topology of mammalian planar cell polarity protein Vangl1.

    abstract::Vangl1 and Vangl2 are membrane proteins that play an important role in neurogenesis, and Vangl1/Vangl2 mutations cause neural tube defects in mice and humans. At the cellular level, Vangl proteins regulate the establishment of planar cell polarity (PCP), a process requiring membrane assembly of asymmetrically distribu...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi101767a

    authors: Iliescu A,Gravel M,Horth C,Apuzzo S,Gros P

    更新日期:2011-03-29 00:00:00

  • RNase T1 variant RV cleaves single-stranded RNA after purines due to specific recognition by the Asn46 side chain amide.

    abstract::Attempts to alter the guanine specificity of ribonuclease T1 (RNase T1) by rational or random mutagenesis have failed so far. The RNase T1 variant RV (Lys41Glu, Tyr42Phe, Asn43Arg, Tyr45Trp, and Glu46Asn) designed by combination of a random and a rational mutagenesis approach, however, exhibits a stronger preference t...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi035961f

    authors: Czaja R,Struhalla M,Höschler K,Saenger W,Sträter N,Hahn U

    更新日期:2004-03-16 00:00:00

  • Carbon monoxide dehydrogenase from Clostridium thermoaceticum: quaternary structure, stoichiometry of its SDS-induced dissociation, and characterization of the faster-migrating form.

    abstract::The molecular mass (M(r)) of the nickel- and iron-sulfur-containing enzyme CO dehydrogenase from Clostridium thermoaceticum was determined by sedimentation equilibrium ultracentrifugation to be 300,000 +/- 30,000 Da. Since the enzyme is known to contain equal numbers of two types of subunits (M(r) = 82,000 Da for alph...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi9511853

    authors: Xia J,Sinclair JF,Baldwin TO,Lindahl PA

    更新日期:1996-02-13 00:00:00