Purification and properties of nuclease SP.


:Single-strand-specific nucleases are a diverse and important group of enzymes that are able to cleave a variety of DNA structures present in duplex molecules. Nuclease SP, an enzyme from spinach, has been purified to apparent homogeneity, allowing for the unambiguous characterization of a number of its physical properties as well as its DNA strand cleavage specificities. The effects of ionic strength, pH, divalent metal cations, and temperature on nuclease SP activity have been examined in detail. Nuclease SP was found to be quite thermostable and could be stimulated by Co2+. In addition, the cleavage of UV-damaged and undamaged supercoiled plasmid substrates under a variety of conditions suggests that at least two types of structures are recognized and processed by nuclease SP: UV photoproduct-induced distortions and unwound "nuclease hypersensitive sites". These studies indicate that nuclease SP is functionally related to other single-strand-specific nucleases and is a potential enzymatic tool for probing and manipulating various types of DNA structures.






Strickland JA,Marzilli LG,Puckett JM Jr,Doetsch PW




Has Abstract


1991-10-08 00:00:00












  • 31P NMR studies of ATP synthesis and hydrolysis kinetics in the intact myocardium.

    abstract::The origin of the nuclear magnetic resonance (NMR)-measurable ATP in equilibrium Pi exchange and whether it can be used to determine net oxidative ATP synthesis rates in the intact myocardium were examined by detailed measurements of ATP in equilibrium Pi exchange rates in both directions as a function of the myocardi...


    pub_type: 杂志文章


    authors: Kingsley-Hickman PB,Sako EY,Mohanakrishnan P,Robitaille PM,From AH,Foker JE,Uğurbil K

    更新日期:1987-11-17 00:00:00

  • Spectroscopic and kinetic studies of lipases solubilized in reverse micelles.

    abstract::The conformation and activity of three different lipases have been studied in reverse micelles formed by sodium bis(2-ethylhexyl) sulfosuccinate (AOT) in isooctane. In the case of human pancreatic lipase, the conformation of the polypeptide chain--as judged from far-UV circular dichroism measurements--is only slightly...


    pub_type: 杂志文章


    authors: Walde P,Han D,Luisi PL

    更新日期:1993-04-20 00:00:00

  • Role of the cochaperone Tpr2 in Hsp90 chaperoning.

    abstract::The molecular chaperones Hsp90 and Hsp70 are highly regulated by various cochaperones that participate in the activation of steroid receptors. Here we study Tpr2 (also called DjC7), a TPR domain-containing type III J protein implicated in steroid receptor chaperoning. We propose that Tpr2 plays a role in the Hsp90-dep...


    pub_type: 杂志文章


    authors: Moffatt NS,Bruinsma E,Uhl C,Obermann WM,Toft D

    更新日期:2008-08-05 00:00:00

  • Trifunctional dinuclear platinum complexes as DNA-protein cross-linking agents.

    abstract::The trifunctional dinuclear platinum compounds 1,2/c,c [[cis-PtCl(NH(3))(2)]mu-H(2)N(CH(2))(6)NH(2)[cis-PtCl(2)(NH(3))]](+) and 1,2/t,c [[trans-PtCl(NH(3))(2)]mu-H(2)N(CH(2))(6)NH(2)[cis-PtCl(2)(NH(3))]](+) contain a monofunctional platinum coordination sphere linked to a cis-[PtCl(2)(amine)(2)] moiety. The compounds ...


    pub_type: 杂志文章


    authors: Kloster M,Kostrhunova H,Zaludova R,Malina J,Kasparkova J,Brabec V,Farrell N

    更新日期:2004-06-22 00:00:00

  • Membrane-binding amphipathic alpha-helical peptide derived from CTP:phosphocholine cytidylyltransferase.

    abstract::A peptide corresponding to a portion of the amphipathic alpha-helical region of CTP:phosphocholine cytidylyltransferase was synthesized. This region of the enzyme was proposed to be the membrane-binding domain [Kalmar, G.B., Kay, R.J., Lachance, A., Aebersold, R., & Cornell, R.B. (1990) Proc. Natl. Acad. Sci. U.S.A. 8...


    pub_type: 杂志文章


    authors: Johnson JE,Cornell RB

    更新日期:1994-04-12 00:00:00

  • "Nonclassical" secretion of annexin A2 to the lumenal side of the enterocyte brush border membrane.

    abstract::Annexin A2 is a member of the annexin family of Ca(2+)-dependent lipid binding proteins and believed to be engaged in membrane transport processes in a number of cell types. In small intestinal enterocytes, we localized annexin A2 to the brush border region, where it was found mainly on the lumenal side of the microvi...


    pub_type: 杂志文章


    authors: Danielsen EM,van Deurs B,Hansen GH

    更新日期:2003-12-16 00:00:00

  • Structure-function analysis of the mammalian DNA polymerase beta active site: role of aspartic acid 256, arginine 254, and arginine 258 in nucleotidyl transfer.

    abstract::The crystal structure of the catalytic domain of rat DNA polymerase beta revealed that Asp256 is located in proximity to the previously identified active site residues Asp190 and Asp192. We have prepared and kinetically characterized the nucleotidyl transfer activity of wild type and several mutant forms of human and ...


    pub_type: 杂志文章


    authors: Menge KL,Hostomsky Z,Nodes BR,Hudson GO,Rahmati S,Moomaw EW,Almassy RJ,Hostomska Z

    更新日期:1995-12-12 00:00:00

  • Multiple forms of phosphodeoxyribomutase from Escherichia coli. Physical and chemical characterization.

    abstract::Phosphodeoxyribomutase from Escherichia coli has been purified to homogeneity. Chromatography on DEAE-Sephadex revealed three peaks of enzyme activity, designated form I, form II, and form III. Form III could be further separated into form III-1 and form III-2 by polyacrylamide gel electrophoresis. The four different ...


    pub_type: 杂志文章


    authors: Leer JC,Hammer-Jespersen K

    更新日期:1975-02-11 00:00:00

  • Lipidic sponge phase crystal structure of a photosynthetic reaction center reveals lipids on the protein surface.

    abstract::Membrane proteins are embedded in a lipid bilayer and maintain strong interactions with lipid molecules. Tightly bound lipids are responsible for vertical positioning and integration of proteins in the membrane and for assembly of multisubunit complexes and occasionally act as substrates. In this work we present the l...


    pub_type: 杂志文章


    authors: Wöhri AB,Wahlgren WY,Malmerberg E,Johansson LC,Neutze R,Katona G

    更新日期:2009-10-20 00:00:00

  • Combinatorial optimization of the DNA cleaving Ni(II) x Xaa-Xaa-His metallotripeptide domain.

    abstract::A positional-scanning combinatorial protocol was employed to optimize the deoxyribose-based cleavage of B-form DNA by Ni(II) x Xaa-Xaa-His metallopeptides. This procedure employed 18 naturally occurring amino acids (excluding Cys and Trp) to generate two libraries in which the first and second positions of the peptide...


    pub_type: 杂志文章


    authors: Huang X,Pieczko ME,Long EC

    更新日期:1999-02-16 00:00:00

  • DNA degradation by bleomycin: evidence for 2'R-proton abstraction and for C-O bond cleavage accompanying base propenal formation.

    abstract::Reaction of poly(dA-[2'S-3H]dU) with activated bleomycin yields [3H]uracil propenal that completely retains the tritium label. In contrast, we have previously shown that reaction of poly(dA-[2'R-3H]dU) with activated bleomycin affords unlabeled uracil propenal [Wu, J. C., Kozarich, J. W., & Stubbe, J. (1983) J. Biol. ...


    pub_type: 杂志文章


    authors: Ajmera S,Wu JC,Worth L Jr,Rabow LE,Stubbe J,Kozarich JW

    更新日期:1986-10-21 00:00:00

  • Crystal structure of rat apo-heme oxygenase-1 (HO-1): mechanism of heme binding in HO-1 inferred from structural comparison of the apo and heme complex forms.

    abstract::Heme oxygenase (HO) catalyzes the oxidative cleavage of heme to biliverdin by utilizing O(2) and NADPH. HO (apoHO) was crystallized as twinned P3(2) with three molecules per asymmetric unit, and its crystal structure was determined at 2.55 A resolution. Structural comparison of apoHO and its complex with heme (HO-heme...


    pub_type: 杂志文章


    authors: Sugishima M,Sakamoto H,Kakuta Y,Omata Y,Hayashi S,Noguchi M,Fukuyama K

    更新日期:2002-06-11 00:00:00

  • Cysteine-rich secretory protein 3 is a ligand of alpha1B-glycoprotein in human plasma.

    abstract::Human cysteine-rich secretory protein 3 (CRISP-3; also known as SGP28) belongs to a family of closely related proteins found in mammals and reptiles. Some mammalian CRISPs are known to be involved in the process of reproduction, whereas some of the CRISPs from reptiles are neurotoxin-like substances found in lizard sa...


    pub_type: 杂志文章


    authors: Udby L,Sørensen OE,Pass J,Johnsen AH,Behrendt N,Borregaard N,Kjeldsen L

    更新日期:2004-10-12 00:00:00

  • Photooxidation pathway of chlorophyll Z in photosystem II as Studied by Fourier transform infrared spectroscopy.

    abstract::The oxidation pathway of chlorophyll Z (ChlZ) in photosystem II (PSII) at cryogenic temperatures was studied by means of light-induced Fourier transform infrared (FTIR) difference spectroscopy. To examine the involvement of redox-active beta-carotene (Car) in the pathway, two Car molecules in Mn-depleted PSII membrane...


    pub_type: 杂志文章


    authors: Kitajima Y,Noguchi T

    更新日期:2006-02-14 00:00:00

  • Lipoprotein lipase catalyzed hydrolysis of water-soluble p-nitrophenyl esters. Inhibition by apolipoprotein C-II.

    abstract::Bovine milk lipoprotein lipase (LpL) catalyzes the hydrolysis of the water-soluble esters p-nitrophenyl acetate (PNPA) and p-nitrophenyl butyrate (PNPB). The same protein and same active site are involved in hydrolysis of water-soluble p-nitrophenyl esters and emulsified trioleoylglycerol since (a) trioleoylglycerol h...


    pub_type: 杂志文章


    authors: Quinn DM,Shirai K,Jackson RL,Harmony JA

    更新日期:1982-12-21 00:00:00

  • DNA minor groove adducts formed by a platinum-acridine conjugate inhibit association of tata-binding protein with its cognate sequence.

    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...


    pub_type: 杂志文章


    authors: Budiman ME,Bierbach U,Alexander RW

    更新日期:2005-08-23 00:00:00

  • Glycosylated polyproline II rods with kinks as a structural motif in plant hydroxyproline-rich glycoproteins.

    abstract::Hydroxyproline-rich glycoproteins (HRGPs) are the major proteinaceous components of higher plant walls and the predominant components of the cell wall of the green alga Chlamydomonas reinhardtii. The GP1 protein, an HRGP of the C. reinhardtii wall, is shown to adopt a polyproline II helical configuration and to carry ...


    pub_type: 杂志文章


    authors: Ferris PJ,Woessner JP,Waffenschmidt S,Kilz S,Drees J,Goodenough UW

    更新日期:2001-03-06 00:00:00

  • Kinetic characterization of recombinant human acidic mammalian chitinase.

    abstract::Human acidic mammalian chitinase (AMCase), a member of the family 18 glycosyl hydrolases, is one of the important proteins involved in Th2-mediated inflammation and has been implicated in asthma and allergic diseases. Inhibition of AMCase results in decreased airway inflammation and airway hyper-responsiveness in a mo...


    pub_type: 杂志文章


    authors: Chou YT,Yao S,Czerwinski R,Fleming M,Krykbaev R,Xuan D,Zhou H,Brooks J,Fitz L,Strand J,Presman E,Lin L,Aulabaugh A,Huang X

    更新日期:2006-04-11 00:00:00

  • Structure of Helicobacter pylori catalase, with and without formic acid bound, at 1.6 A resolution.

    abstract::Helicobacter pylori produces one monofunctional catalase, encoded by katA (hp0875). The crystal structure of H. pylori catalase (HPC) has been determined and refined at 1.6 A with crystallographic agreement factors R and R(free) of 17.4 and 21.9%, respectively. The crystal exhibits P2(1)2(1)2 space group symmetry and ...


    pub_type: 杂志文章


    authors: Loewen PC,Carpena X,Rovira C,Ivancich A,Perez-Luque R,Haas R,Odenbreit S,Nicholls P,Fita I

    更新日期:2004-03-23 00:00:00

  • Thermodynamics of nucleotide and inhibitor binding to wild-type and ispinesib-resistant forms of human kinesin spindle protein.

    abstract::Current antimitotic cancer chemotherapy based on vinca alkaloids and taxanes target tubulin, a protein required not only for mitotic spindle formation but also for the overall structural integrity of terminally differentiated cells. Among many innovations targeting specific mitotic events, inhibition of motor enzymes ...


    pub_type: 杂志文章


    authors: Sheth PR,Basso A,Duca JS,Lesburg CA,Ogas P,Gray K,Nale L,Mannarino AF,Prongay AJ,Le HV

    更新日期:2009-11-24 00:00:00

  • Probing the heme axial ligation in the CO-sensing CooA protein with magnetic circular dichroism spectroscopy.

    abstract::The combination of UV/visible/near-IR variable-temperature magnetic circular dichroism (VTMCD) and EPR spectroscopies has been used to investigate the spin states and axial ligation of the heme group in oxidized, reduced, and CO-bound reduced forms of the Rhodospirillum rubrum CO oxidation transcriptional activator pr...


    pub_type: 杂志文章


    authors: Dhawan IK,Shelver D,Thorsteinsson MV,Roberts GP,Johnson MK

    更新日期:1999-09-28 00:00:00

  • Interactions of soluble penicillin-binding protein 2a of methicillin-resistant Staphylococcus aureus with moenomycin.

    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...


    pub_type: 杂志文章


    authors: Graves-Woodward K,Pratt RF

    更新日期:1999-08-10 00:00:00

  • Rapid changes in the phosphoproteome show diverse cellular responses following stimulation of human lung fibroblasts with endothelin-1.

    abstract::The rapid phosphorylation and dephosphorylation of a variety of proteins downstream of the endothelin receptors A and B was investigated following stimulation of human lung fibroblasts with endothelin-1. Changes in the phosphorylation of proteins involved in the cell cycle, cytoskeleton, membrane channels, transcripti...


    pub_type: 杂志文章


    authors: Stannard C,Soskic V,Godovac-Zimmermann J

    更新日期:2003-12-02 00:00:00

  • Structural, Functional, and Dynamical Responses of a Protein in a Restricted Environment Imposed by Macromolecular Crowding.

    abstract::The intercellular environment is known to be very different from the environment where most of the elementary biological processes are studied in the laboratory. As a result, there was a considerable effort on cell mimicking either by confinement or by introducing macromolecular crowding. In the present study, dextran...


    pub_type: 杂志文章


    authors: Das N,Sen P

    更新日期:2018-10-23 00:00:00

  • New insights into inhibitor design from the crystal structure and NMR studies of Escherichia coli GAR transformylase in complex with beta-GAR and 10-formyl-5,8,10-trideazafolic acid.

    abstract::The crystal structure of Escherichia coli GAR Tfase at 2.1 A resolution in complex with 10-formyl-5,8,10-trideazafolic acid (10-formyl-TDAF, K(i) = 260 nM), an inhibitor designed to form an enzyme-assembled multisubstrate adduct with the substrate, beta-GAR, was studied to determine the exact nature of its inhibitory ...


    pub_type: 杂志文章


    authors: Greasley SE,Yamashita MM,Cai H,Benkovic SJ,Boger DL,Wilson IA

    更新日期:1999-12-21 00:00:00

  • Thermodynamics of reversible monomer-dimer association of tubulin.

    abstract::The equilibrium between the rat brain tubulin alpha beta dimer and the dissociated alpha and beta monomers has been studied by analytical ultracentrifugation with use of a new method employing short solution columns, allowing rapid equilibration and hence short runs, minimizing tubulin decay. Simultaneous analysis of ...


    pub_type: 杂志文章


    authors: Sackett DL,Lippoldt RE

    更新日期:1991-04-09 00:00:00

  • Chaotropic agents and increased matrix volume enhance binding of mitochondrial cyclophilin to the inner mitochondrial membrane and sensitize the mitochondrial permeability transition to [Ca2+].

    abstract::Binding of mitochondrial cyclophilin (CyP) to the inner mitochondrial membrane is induced by treatment of mitochondria with thiol reagents or oxidative stress and correlates with a sensitization to [Ca2+] of the cyclosporin A-sensitive mitochondrial permeability transition pore (MTP) [Connern, C. P., & Halestrap, A. P...


    pub_type: 杂志文章


    authors: Connern CP,Halestrap AP

    更新日期:1996-06-25 00:00:00

  • Characterization of the quaternary structure and conformational properties of the human stem cell inhibitor protein LD78 in solution.

    abstract::The human LD78 protein (sometimes referred to as human macrophage inflammatory protein-1 alpha) has been shown to protect multipotential hemopoietic stem cells from the effects of cytotoxic agents. Administration of the recombinant stem cell inhibitor molecule LD78 as an adjunct to chemotherapy has potential clinical ...


    pub_type: 杂志文章


    authors: Patel SR,Evans S,Dunne K,Knight GC,Morgan PJ,Varley PG,Craig S

    更新日期:1993-05-25 00:00:00

  • Discovery of a novel L-lyxonate degradation pathway in Pseudomonas aeruginosa PAO1.

    abstract::The l-lyxonate dehydratase (LyxD) in vitro enzymatic activity and in vivo metabolic function were assigned to members of an isofunctional family within the mandelate racemase (MR) subgroup of the enolase superfamily. This study combined in vitro and in vivo data to confirm that the dehydration of l-lyxonate is the bio...


    pub_type: 杂志文章


    authors: Ghasempur S,Eswaramoorthy S,Hillerich BS,Seidel RD,Swaminathan S,Almo SC,Gerlt JA

    更新日期:2014-05-27 00:00:00

  • Adenosine cyclic 3',5'-monophosphate dependent protein kinase: active site directed inhibition by Cibacron Blue F3GA.

    abstract::Cibacron Blue F3GA, the polycyclic blue chromophore of Blue Dextran 2000, inhibits the catalytic subunit of bovine brain protein kinase. The rate of inactivation exhibits a hyperbolic dependence on the dye concentration. This suggests that an enzyme-dye complex forms prior to inactivation. Protein and peptide substrat...


    pub_type: 杂志文章


    authors: Witt JJ,Roskoski R Jr

    更新日期:1980-01-08 00:00:00