Abstract:
:Heparanase is a beta-D-endoglucuronidase that cleaves heparan sulfate (HS) and has been implicated in many important physiological and pathological processes, including tumor cell metastasis, angiogenesis, and leukocyte migration. We report herein the identification of active-site residues of human heparanase. Using PSI-BLAST and PHI-BLAST searches of sequence databases, similarities were identified between heparanase and members of several of the glycosyl hydrolase families (10, 39, and 51) from glycosyl hydrolase clan A (GH-A), including strong local identities to regions containing the critical active-site catalytic proton donor and nucleophile residues that are conserved in this clan of enzymes. Furthermore, secondary structure predictions suggested that heparanase is likely to contain an (alpha/beta)(8) TIM-barrel fold, which is common to the GH-A families. On the basis of sequence alignments with a number of glycosyl hydrolases from GH-A, Glu(225) and Glu(343) of human heparanase were identified as the likely proton donor and nucleophile residues, respectively. The substitution of these residues with alanine and the subsequent expression of the mutant heparanases in COS-7 cells demonstrated that the HS-degrading capacity of both was abolished. In contrast, the alanine substitution of two other glutamic acid residues (Glu(378) and Glu(396)), both predicted to be outside the active site, did not affect heparanase activity. These data suggest that heparanase is a member of the clan A glycosyl hydrolases and has a common catalytic mechanism that involves two conserved acidic residues, a putative proton donor at Glu(225) and a nucleophile at Glu(343).
journal_name
Biochemistryjournal_title
Biochemistryauthors
Hulett MD,Hornby JR,Ohms SJ,Zuegg J,Freeman C,Gready JE,Parish CRdoi
10.1021/bi002080psubject
Has Abstractpub_date
2000-12-26 00:00:00pages
15659-67issue
51eissn
0006-2960issn
1520-4995pii
bi002080pjournal_volume
39pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::Conformational states of the Na,K-ATPase and rates of transition between these are studied using the fluorescent dye eosin as a marker for the Na+ form (E1) and occlusion of 86Rb+ as a marker for the K+ form (E2). The aim of the present paper is to propose that the E1 form of the Na,K-ATPase can be liganded with a num...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00194a022
更新日期:1994-07-19 00:00:00
abstract::Solvent reorganization can contribute significantly to the energetics of protein-protein interactions. However, our knowledge of the magnitude of the energetic contribution is limited, in part, by a dearth of quantitative experimental measurements. The biotin repressor forms a homodimer as a prerequisite to DNA bindin...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi400952m
更新日期:2013-09-24 00:00:00
abstract::The instantaneous electrochromic response of carotenoids associated with the B800-850 light-harvesting complex of Rhodobacter sphaeroides has been used widely as an intrinsic probe of membrane potential. In the present study, the structural basis for this phenomenon was examined by phospholipase A2 digestion of chroma...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00054a017
更新日期:1993-01-26 00:00:00
abstract::The Wiskott-Aldrich syndrome protein (WASP) and neural WASP (N-WASP) are key players in regulating actin cytoskeleton via the Arp2/3 complex. It has been widely reported that the WASP proteins are activated by Rho family small GTPase Cdc42 and that Rac1 acts through SCAR/WAVE proteins. However, a systematic study of t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi062152y
更新日期:2007-03-20 00:00:00
abstract::The role of the protein shell in the formation of the hydrous ferric oxide core of ferritin is poorly understood. A VO2+ spin probe study was undertaken to characterize the initial complex of Fe2+ with horse spleen apoferritin (96% L-subunits). A competitive binding study of VO2+ and Fe2+ showed that the two metals co...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00102a012
更新日期:1991-09-24 00:00:00
abstract::Leucine aminopeptidase (LAP) is a protease requiring two divalent metal cations per subunit for activity. Zn2+, Mg2+, and Co2+ metal-substituted forms of LAP have been prepared and investigated kinetically. Substitution of metal into the two binding sites independently resulted in the preparation of Zn2+Zn2+, Mg2+Zn2+...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00285a010
更新日期:1983-08-02 00:00:00
abstract::Continuum methods were used to calculate the electrostatic contributions of charged and polar side chains to the overall stability of a small 41-residue helical protein, the peripheral subunit-binding domain. The results of these calculations suggest several residues that are destabilizing, relative to hydrophobic iso...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi992091m
更新日期:2000-02-08 00:00:00
abstract::By exposing photosystem II (PSII) samples to an incrementing number of excitation flashes at room temperature, followed by freezing, we could compare the Mn-derived multiline EPR signal from the S2 oxidation state as prepared by 1, 5, 10, and 25 flashes of light. While the S2 multiline signals exhibited by these sampl...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi026848c
更新日期:2003-03-11 00:00:00
abstract::Heparan sulfate (HS) proteoglycans (PGs) interact with a number of extracellular signaling proteins, thereby playing an essential role in the regulation of many physiological processes. These interactions are important for both normal signal transduction and regulation of the tissue distribution of signaling molecules...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi6025424
更新日期:2007-04-03 00:00:00
abstract::Sucrose density gradient shift assays were used to study the interactions of human glucocorticoid receptors (GR) with small DNA fragments either containing or lacking glucocorticoid response element (GRE) DNA consensus sequences. When crude cytoplasmic extracts containing [3H]triamcinolone acetonide [( 3H]TA) labeled ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00431a003
更新日期:1989-03-07 00:00:00
abstract::Inhibition of the yeast F(0)F(1)-ATP synthase by the regulatory peptides IF1 and STF1 was studied using intact mitochondria and submitochondrial particles from wild-type cells or from mutants lacking one or both peptides. In intact mitochondria, endogenous IF1 only inhibited uncoupled ATP hydrolysis and endogenous STF...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi034394t
更新日期:2003-06-24 00:00:00
abstract::Sarcosine oxidase from Corynebacterium sp. P-1 binds 2 mol of tetrahydrofolate/mol of enzyme (KD = 8.8 microM). The same stoichiometry is observed with tetrahydropteroyltetraglutamate (KD = 15.4 microM). Binding is also observed with pteroyltetraglutamate and with 5-formyltetrahydrofolate. In the case of the pteroylmo...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00397a029
更新日期:1987-11-17 00:00:00
abstract::Ferrochelatase is the terminal enzyme in the heme biosynthetic pathway. It catalyzes the insertion of ferrous iron into protoporphyrin IX to produce protoheme IX. The crystal structures of ferrochelatase from Saccharomyces cerevisiae in free form, in complex with Co(II), a substrate metal ion, and in complex with two ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0260785
更新日期:2002-11-19 00:00:00
abstract::The folding of the peptide chain of the bovine heart oxoglutarate carrier in the inner mitochondrial membrane and in the membrane of reconstituted proteoliposomes has been investigated by enzymatic and immunochemical approaches using proteinase K and polyclonal site-directed antibodies, respectively. Two peptides corr...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00178a030
更新日期:1994-03-29 00:00:00
abstract::The expression of apoE mRNA in the adrenal gland is inversely correlated to steroidogenesis and directly correlated to the level of cholesteryl ester stores. To further investigate the relationship between apoE and cellular cholesterol homeostasis, several parameters of cholesterol metabolism in the murine Y1 adrenal ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00183a007
更新日期:1994-05-03 00:00:00
abstract::The beta- and delta-subunits of the nicotinic acetylcholine receptor from Torpedo californica were covalently photolabeled at the synaptic surface with the ATP photoaffinity analogue [alpha-32P]-8-azido-ATP. The specificity of labeling for nucleotide binding sites was demonstrated by the saturation of labeling with in...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00080a002
更新日期:1993-07-27 00:00:00
abstract::For some hormone receptors, the early events of signal transduction depend on their molecular arrangement and interactions at the cell surface. An understanding of the mechanism of signal transduction in general needs a careful analysis of the receptor distribution. Here, we present the first quantitative measurement ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00124a004
更新日期:1992-03-10 00:00:00
abstract::Remodeling of photosynthetic machinery induced by growing spinach plants under low light intensities reveals an up-regulation of light-harvesting complexes and down-regulation of photosystem II and cytochrome b6f complexes in intact thylakoids and isolated grana membranes. The antenna size of PSII increased by 40-60% ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi700748y
更新日期:2007-10-02 00:00:00
abstract::We have investigated the effect of increasing percentages of methylated pyrimidines on the structure of poly(dA-dU).poly(dA-dU). This was done by synthesizing analogous polynucleotides that contained deoxythymidine residues as well as deoxyuridine residues and observing their 31P NMR spectra in increasing amounts of C...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00341a033
更新日期:1985-09-24 00:00:00
abstract::Glu58 is known to participate in phosphodiester transesterification catalyzed by the enzyme RNase T1. For Glu58 RNase T1, an altered mechanism has been proposed in which His40 replaces Glu58 as the base catalyst [Steyaert, J., Hallenga, K., Wyns, L., & Stanssens, P. (1990) Biochemistry 29, 9064-9072]. Glu58Ala Rnase T...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00173a006
更新日期:1994-02-22 00:00:00
abstract::Telomerase is a ribonucleoprotein that participates in the maintenance of telomere length. Its activity is up-regulated in many tumor types, suggesting that it may be a novel target for chemotherapy. The RNA component of telomerase contains an active site that plays at least two roles&sbd;binding telomere ends and tem...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi970438k
更新日期:1997-09-30 00:00:00
abstract::We have used diethyl pyrocarbonate (DEP), which carbethoxylates adenine bases, and dimethyl sulfate (DMS), which methylates guanine residues and single-stranded cytosines, to probe bases in open complexes between RNA polymerase and the lac UV5 promoter in vitro. We compared the kinetics of reactivity between bases in ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00436a040
更新日期:1989-05-16 00:00:00
abstract::The inactivation of cytochrome P450 2B4 by aldehydes in a reconstituted enzyme system requires molecular oxygen and NADPH and is not prevented by the addition of catalase, superoxide dismutase, epoxide hydrolase, glutathione, or ascorbic acid. A strong correlation between loss of enzymatic activity and bleaching of th...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9630568
更新日期:1997-04-22 00:00:00
abstract::A 19-amino acid residue peptide, Gly-Trp-Leu-Lys-Ile-Lys-Ala-Ala-Met-Arg-Trp-Gly-Phe-Phe-Val-Arg-Lys-Lys- Ala, corresponding to the basic amphiphilic alpha-helix (BAA) motif at the C-terminus of a recombinant tobacco calmodulin-binding protein, TCB60, was synthesized. The interaction of the synthetic binding domain wi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9621674
更新日期:1997-02-25 00:00:00
abstract::This paper reports on NMR-molecular mechanics structural studies of the (-)-trans-anti-[MC]dG adduct positioned opposite dC in the sequence context of the d(C1-C2-A3-T4-C5-[MC]G6-C7-T8-A9-C10-C11).d(G12-G13-T14++ +-A15-G16-C17-G18- A19-T20-G21-G22) duplex [designated (-)-trans-anti-[MC]dG.dC 11-mer duplex]. This adduc...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00018a029
更新日期:1995-05-09 00:00:00
abstract::The inactivation of excited rhodopsin in the presence of ATP, rhodopsin kinase, and/or arrestin has been studied from its effect on the two subsequent steps in the light-induced enzymatic cascade: metarhodopsin II catalyzed activation of G-protein and G-protein-dependent activation of cGMP phosphodiesterase. The inact...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00405a049
更新日期:1988-03-08 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
abstract::The characteristic period four oscillation patterns of oxygen evolution induced by a train of single-turnover flashes were measured as a function of temperature in dark-adapted photosystem II (PS II) membrane fragments that were reconstituted with native plastoquinone-9 (PQ-9) by a recently developed procedure [Kurrec...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi962302c
更新日期:1997-03-04 00:00:00
abstract::Although the short-chain dehydrogenase/reductase (SDR) superfamily contains a very large number of members defined in annotated databases and by biochemical and structural studies, very few SDR enzymes have been identified that have a homologous partner catalyzing the same reaction but with an opposite stereospecifici...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0118005
更新日期:2002-02-26 00:00:00
abstract::The tyrosine kinase c-Abl is implicated in a variety of cellular processes that are tightly regulated by c-Abl kinase activity and/or by interactions between c-Abl and other signaling molecules. The interaction of c-Abl with the Abl interactor protein Abi2 is shown to be negatively regulated by phosphorylation of seri...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi701533j
更新日期:2008-01-22 00:00:00