ライフサイエンスコーパス: heparinase

1 which was reversed by antifibrinolytics and heparinase.

2 to treatment of macrophages with trypsin or heparinase.

3 PG and in HSPG-expressing cells treated with heparinase.

4 sensitive to both chondroitin ABC lyase and heparinase.

5 main, and by incubating the blastocysts with heparinase.

6 excess heparan sulfate and by treatment with heparinase.

7 sion was reversed by tetrahydrolipstatin and heparinase.

8 y soluble heparin and by treating cells with heparinase.

9 rin or by the pretreatment of the cells with heparinase.

10 broadest known substrate specificity of the heparinases.

11 bolished when CHO-K1 cells were treated with heparinases.

12 ycosaminoglycans (GAGs) on the action of the heparinases.

13 age of entry was observed in CF treated with heparinases.

14 , however, contained three, compared to two, heparinase 1-resistant sequences separated by larger con

15 Cellular treatment with heparinase and chondroitinase ABC inhibited expression b

16 Pretreatment of cells with heparinase and heparitinase abolished more than 80% of t

17 ell surface heparan sulfate proteoglycans by heparinase and heparitinase but not by treatment with co

18 with the glycosaminoglycan-degrading enzymes heparinase and heparitinase suggesting the specific invo

19 was examined with or without the addition of heparinase and heparitinase to cell incubation mixtures.

20 Heparinase and heparitinase treatment of cells reduced b

21 lude inhibition of FPX cleavage by bacterial heparinase and mammalian heparanase enzymes with inhibit

22 rin formation) greater than 1 minute between heparinase and standard thrombelastogram (TEG) is associ

23 Among others, these enzymes include the heparinases and an unusual glycuronidase that hydrolyzes

24 on of heparin-like glycosaminoglycans by the heparinases and mutant heparinases could pave the way to

25 By disrupting (with sulfation inhibitors and heparinase) and partially reconstituting (with heparin)

26 ls and primary neurons by heparin, chlorate, heparinase, and genetic knockdown of a key HSPG syntheti

27 Heparinases are bacterial enzymes that are powerful tool

28 -like glycosaminoglycan degrading enzymes or heparinases are powerful tools that have enabled the elu

29 of porcine intestinal heparin with bacterial heparinase), as well as a heparin-derived pharmaceutical

30 fate from the cell surface by treatment with heparinase blocked HCV attachment.

31 Importantly, treatment with heparin or heparinase blocked LRP1-mediated cellular uptake of Abet

32 anced uptake was reduced by more than 80% by heparinase but was unaffected by the 39-kDa protein.

33 lost when intact cells were predigested with heparinases but not chondroitinases.

34 The sensor is sensitive to the inhibition of heparinase by OSCS until a concentration of 200 pg/ml re

35 saccharides obtained through the controlled, heparinase-catalyzed depolymerization of heparin.

36 cess heparin or pretreatment of acini with a heparinase cocktail each inhibited Ad5 transduction by a

37 minating heparan sulfate proteoglycans using heparinase completely abrogated the mechanical induction

38 g distinctions in substrate specificities of heparinases could be used to isolate oligosaccharides wi

39 osaminoglycans by the heparinases and mutant heparinases could pave the way to the development of muc

40 independent means of disrupting syndecan-4: heparinase degradation of HS glycosaminoglycans or suppr

41 These proteoglycans were sensitive to heparinase, demonstrating that K562 cells synthesize bFG

42 The three heparinases derived from Flavobacterium heparinum are po

43 utility of this enzyme in the sequencing of heparinase-derived HSGAG oligosaccharides.

44 increased binding of both proteins, whereas heparinase digestion and competition with heparin/HS inh

45 pressing activity of EHS-BM was sensitive to heparinase digestion but not to chondroitinase ABC or hy

46 assay to assess the purity of heparin using heparinase digestion followed by size-exclusion HPLC ana

47 Heparinase digestion of the endothelial cell's glycocaly

48 HIP was not released by heparinase digestion suggesting that the association is

49 ular epithelial cells and can be released by heparinase digestion.

50 Furthermore, treatment of cells with heparinase diminished infection with SBV, confirming tha

51 tease (EC 3.4.21.16); (iv) lyase activity of heparinase (EC 4.1.1.7); and (v) ligase activity of pyru

52 end of oligosaccharides resulting from prior heparinase eliminative cleavage.

53 We recently described the P. aeruginosa heparinase-encoding gene, hepP, whose expression was sig

54 zation, while pretreatment of HIS cells with heparinase enzyme or with anti-3-OS HS (G2) peptide sign

55 chain, which it protects from degradation by heparinase enzymes.

56 Finally, pretreatment of cells with heparinase failed to abolish apoE inhibition of smooth m

57 Heparinase III is the unique member of the heparinase family of heparin-degrading lyases that recog

58 The heparinases from Flavobacterium heparinum are lyases tha

59 The heparinases from Flavobacterium heparinum are powerful t

60 ools used for the production of LMWH are the heparinases from Flavobacterium heparinum, specifically

61 , prostaglandin-endoperoxide synthase-2, and heparinase genes.

62 polymerization using the bacterially derived heparinases, given the structural understanding of their

63 Heparinase had no effect on adhesion to the peptide.

64 noglycans and treatment of target cells with heparinase had no significant inhibition on cytoadherenc

65 In addition, heparinases have significant therapeutic applications.

66 Heparinase/heparinitase treatment of the basolateral cel

67 Cleavage of HS by heparinase, heparitinase, or heparanase severely reduced

68 Treatment of matrix with heparinase/heparitinase (1 U/ml each) increased LDL bind

69 cysteine-135 as an important amino acid for heparinase I (EC 4.2.2.7) activity.

70 entified the primary heparin binding site of heparinase I (EC 4.2.2.7).

71 t, but did not change the product profile of heparinase I action on heparin; conversely, mutations in

72 w as 10 microM DEPC results in a 85% loss of heparinase I activity in 15 min.

73 Heparinase I activity is restored following hydroxylamin

74 raphy suggested that mutations in CB-1 alter heparinase I activity primarily through decreasing the e

75 nt, but they had a more pronounced effect on heparinase I activity, suggesting a different role for C

76 f the HB-1 region has a pronounced effect on heparinase I activity.

77 results in only a 2- to 3-fold reduction in heparinase I activity.

78 ng that these residues are not essential for heparinase I activity.

79 ride, we showed that the interaction between heparinase I and calcium was essential for proper functi

80 ave shown that calcium binds specifically to heparinase I and have identified two major calcium-bindi

81 Given that both heparinase I and heparinase II contain catalytically cri

82 parinase III, and TRSB was sensitive to both heparinase I and heparinase III.

83 xperiments to answer the question of whether heparinase I binds to calcium and, if so, which regions

84 , strongly suggests that the inactivation of heparinase I by DEPC is specific for histidine residues.

85 60 nm) and that calcium is able to activate heparinase I catalytically.

86 to further understand the mechanism by which heparinase I cleaves its polymer substrate, we sought to

87 also identified a heparin binding domain in heparinase I containing two positively charged clusters

88 to investigate the sequence of events during heparinase I depolymerization of HLGAGs.

89 stions with individual enzymes revealed that heparinase I did not cleave at GlcNH(3)(+) residues; how

90 harides of known structure were subjected to heparinase I digestion and analyzed.

91 ) that are important for calcium binding and heparinase I enzymatic activity.

92 rinization of the circuit via an immobilized heparinase I filter.

93 Heparinase I from Flavobacterium heparinum has important

94 In a second step, heparinase I has a strong preference for cleaving the sa

95 The wild-type heparinase I has four histidine residues; site-directed

96 y, amino acids located in the active site of heparinase I have been identified and mapped.

97 erve concentration-dependent inactivation of heparinase I in the presence of reversible histidine-mod

98 bonate (DEPC); 0.3 mM DEPC results in 95% of heparinase I inactivation in less than 3 min, and as low

99 We show that heparin binding to heparinase I is independent of calcium (Kd of 60 nm) and

100 In an initial step, heparinase I preferentially cleaves exolytically at the

101 hydryl selective labeling of cysteine 135 of heparinase I protects the lysines of the heparin binding

102 Pretreatment of monocytes with heparin or heparinase I resulted in partial inhibition of cell adhe

103 us to propose a model for calcium binding to heparinase I that includes both CB-1 and CB-2 providing

104 nd competition assays, to map the regions of heparinase I that interact specifically with heparin.

105 (Glu207-Ala219) and CB-2 (Thr373-Arg384), in heparinase I that not only are specifically modified by

106 heparin binding site, may bridge heparin to heparinase I through calcium in a ternary complex during

107 escent calcium analog terbium, we found that heparinase I tightly bound divalent and trivalent cation

108 e E2DeltaHVR1-G or E2-G pseudotypes, whereas heparinase I treatment (8 U/ml) of cells reduced 40% E2-

109 preparations of various molecular weights or heparinase I treatment of cells prevented HPV31b infecti

110 d an outer compartment where the immobilized heparinase I was fluidized separately from the blood cel

111 Heparinase I was immobilized onto agarose beads via cyan

112 Chemical and proteolytic digests of heparinase I were used in direct binding and competition

113 However, H203A inactivates heparinase I while a H203D mutant has residual activity,

114 ations in CB-2 not only altered the kcat for heparinase I, but also resulted in incomplete degradatio

115 larly, pretreatment of eukaryotic cells with heparinase I, but not pretreatment of eukaryotic cells w

116 f a glycosidic linkage imparts resistance to heparinase I, II, and III cleavage.

117 ective digestion with pronase, NaOH/NaBH(4), heparinase I, or low pH nitrous acid showed that each HS

118 bition of granule activity by digestion with heparinase I, the failure of proteolysis of the proteogl

119 ing studies of Woodward's reagent K-modified heparinase I, we identified two putative calcium-binding

120 d GAGs examined were effective inhibitors of heparinase I, with IC(50) values ranging from approximat

121 ide evidence that one of the active sites is heparinase I-like, cleaving at hexosamine-sulfated iduro

122 was abrogated by pretreatment of cells with heparinase I.

123 ition of deoxycholic acid, or treatment with heparinase I.

124 n-like glycosaminoglycan depolymerization by heparinase I.

125 ent role for CB-2 in the enzymatic action of heparinase I.

126 cofactor without altering heparin binding to heparinase I.

127 ssary cofactor, in the enzymatic activity of heparinase I.

128 ve mechanism of depolymerization of HLGAG by heparinase I.

129 ne 203 together form the catalytic domain in heparinase I.

130 microenvironment for the catalytic domain of heparinase I.

131 zation of heparin-like glycosaminoglycans by heparinase I.

132 e data for degradation of polymeric HLGAG by heparinase I.

133 sidues together form the catalytic domain of heparinase I.

134 dine(s) lie(s) in or near the active site of heparinase I.

135 aran sulfate or cells that were treated with heparinase I.

136 ned in the heparin binding site) inactivated heparinase I; however, a H203D mutant retained marginal

137 hexasaccharide model compounds, we show that heparinases I and II, but not heparinase III, cleave the

138 As previously observed for heparinases I and II, substrate protection experiments s

139 from Flavobacterium heparinum, specifically heparinases I and II.

140 to venular endothelial cells; treatment with heparinases I and III significantly reduced this binding

141 Treatment with keratanase (3 joints) or heparinases I, II and III (3 joints) caused no significa

142 letion by chondroitinase ABC, keratanase and heparinases I, II and III in vivo.

143 K5 polysaccharide by digestion with combined heparinases I, II, and III.

144 these results are due to residual HSPG since heparinase (I and III) digestion of chlorate-treated cel

145 Heparinase II (no EC number) is one of three lyases isol

146 Heparinase II activity is restored following hydroxylami

147 indicating that cysteine 348 is required for heparinase II activity toward heparin but is not essenti

148 importance of histidines 238, 451 and 579 in heparinase II activity.

149 out to investigate the role of cysteines in heparinase II activity.

150 nes 238, 451, and 579 as being important for heparinase II activity.

151 , strongly suggests that the inactivation of heparinase II by DEPC is specific for histidine residues

152 We show here that heparinase II cleaves heparin-like glycosaminoglycans en

153 Given that both heparinase I and heparinase II contain catalytically critical histidines,

154 agenesis experiments on the 13 histidines of heparinase II corroborated the chemical modification and

155 n with heparan sulfate was unable to protect heparinase II from DEPC inactivation for either of the s

156 Heparinase II generated DeltaHexA-GlcNH(3)(+)(6S),DeltaH

157 In addition, we show that heparinase II has two distinct active sites and provide

158 to investigate the role of the histidines of heparinase II in catalysis.

159 erve concentration-dependent inactivation of heparinase II in the presence of the reversible histidin

160 was found that one of the three cysteines in heparinase II is surface-accessible and possesses unusua

161 Heparinase II is unique among the heparinases in that it

162 Heparinase II is unique among the three enzymes because

163 However, heparinase II preincubation with heparan sulfate was una

164 Substrate protection experiments show that heparinase II preincubation with heparin followed by the

165 Thus, the action of heparinase II requires O-sulfation, whereas heparinase I

166 cell lines with heparin, sodium chlorate, or heparinase II, demonstrating that heparin sulfate proteo

167 eparin-like glycosaminoglycan degradation by heparinase II, we set out to investigate the role of the

168 zation of heparin-like glycosaminoglycans by heparinase II, which possesses the broadest known substr

169 ysteine 189 are functionally unimportant for heparinase II.

170 not cleave at GlcNH(3)(+) residues; however, heparinases II and III showed selective and distinct act

171 ted completely by pretreatment with 15 mU/mL heparinase III (E.C.4.2.2.8) for 2 hours.

172 , removal of cell-surface heparan sulfate by heparinase III abolished the chemorepulsive response to

173 or cell treatment with chondroitinase ABC or heparinase III abolished the mitogenic effects of MK on

174 heparinase II requires O-sulfation, whereas heparinase III acts only on the corresponding non-sulfat

175 lost upon digestion of tissue sections with heparinase III and chondroitinase ABC.

176 Heparinase III and human heparanase specifically cleaved

177 fate, because this activity was abolished by heparinase III digestion of the cells.

178 95 and histidine 510, as being essential for heparinase III enzymatic activity.

179 Heparinase III is the unique member of the heparinase fa

180 ion of heparin and the hydrolysis of HSPG by heparinase III only partially inhibited hIIA PLA2 bindin

181 Removal of the HSs from the cell surface by heparinase III or by silencing syndecan-3 by siRNA parti

182 Enzymatic treatment by heparinase III or chondroitinase ABC also releases TIMP-

183 , and pretreatment of endothelial cells with heparinase III or protease reduced dengue infectivity by

184 of heparan sulfate proteoglycans (HSPG) with heparinase III prevented infection and BM binding by the

185 Moreover, treatment of HUVECs with heparinase III to specifically eliminate the FGF-2 hepar

186 fen, and enzymatic removal of HS chains with heparinase III treatment as well as by site-directed mut

187 In contrast, heparinase III treatment of control cells, with function

188 Heparinase III treatment of the genital tract significan

189 Depletion of cell surface HSPGs by heparinase III treatment or decreased glycosaminoglycan

190 rat uterus by heparin/heparan sulfate and by heparinase III treatment.

191 ith a heparan sulfate antibody revealed that heparinase III treatments removed a substantial fraction

192 A(2S)-GlcNH(3)(+)(6S) disaccharides, whereas heparinase III yielded only the DeltaHexA-GlcNH(3)(+) un

193 lowing results: (1) treatment with bacterial heparinase III, an enzyme that degrades heparan sulfate

194 nsitive to digestion of cell surface HS with heparinase III, and TRSB was sensitive to both heparinas

195 was abolished after treating the cells with heparinase III, but not after chondroitinase treatment.

196 , we show that heparinases I and II, but not heparinase III, cleave the AT-III binding site, leaving

197 ate, pretreatment of conditioned medium with heparinase III, or growth of cells in sodium chlorate, i

198 treatment with either chondroitinase ABC or heparinase III, suggesting that both chondroitin sulfate

199 by heparin and by treatment of HBEC-5i with heparinase III, suggesting that the endothelial receptor

200 cells that had been treated with the enzyme heparinase III, which degrades the glycosaminoglycan sid

201 We have prepared a library of heparinase III-generated HS oligosaccharides fractionate

202 te linkages, whereas the other is presumably heparinase III-like, cleaving at hexosamine-glucuronate

203 idines, we examined the role of histidine in heparinase III.

204 TRSB was sensitive to both heparinase I and heparinase III.

205 ct was lost posttreatment of the tissue with heparinase III.

206 as completely inhibited by pretreatment with heparinase III.

207 and this release was blocked by heparin and heparinase III.

208 Treatment with heparinase-III and p-nitrophenyl-beta-D-xylopyranoside (

209 Heparinase II is unique among the heparinases in that it has broad substrate requirements

210 ndothelial cells, and binding was blocked by heparinase, indicating that secretoneurin stimulates bin

211 Treatment of cells with exogenous heparinase led to a greater than 4-fold increase in apoE

212 availability of HSPG sites as treatment with heparinase or competitors of glycosaminoglycan chain add

213 Treatment of T-cells with either heparinase or heparitinase abolished nucleosome binding

214 urface GAG chains by treatment of cells with heparinase or heparitinase but not chondroitinase marked

215 ll line was also inhibited by digestion with heparinase or heparitinase but not with chondroitinase A

216 fate GAGs on keratinocytes by treatment with heparinase or heparitinase resulted in an 80-90% reducti

217 dothelial cells with chondroitinase, but not heparinase or heparitinase, diminished endothelial bindi

218 is of cell surface HSPGs with, respectively, heparinase or sodium chlorate abrogated HSC adhesion to

219 L1/L2 virus-like particles, is unaffected by heparinase or trypsin pretreatment of HeLa cells.

220 ited by either prior treatment of cells with heparinases or by HS preparations enriched in 3-OS HS.

221 cosaminoglycans in embryonic stem cells with heparinases or sodium chlorate inhibited differentiation

222 oteoglycans because treatment with chlorate, heparinase, or soluble heparin did not prevent Hep II do

223 Antifibrinolytics and heparinase partially reverse the abnormal clotting patte

224 Parasite development in heparinase-pretreated HepG2 cells is inhibited by 65% wh

225 ant NOx production that was not inhibited by heparinase pretreatment, demonstrating that the cells we

226 Heparinase-pretreatment of LRP-deficient fibroblasts or

227 d plasma samples, which were pretreated with heparinase prior to analysis, had the lowest baseline HI

228 Blocking FGF1 receptors with heparinase reduced this effect.

229 teract with a widely expressed, trypsin- and heparinase-resistant cell surface molecule and facilitat

230 Pretreatment of cell cultures with heparinase resulted in reduced bFGF binding to the cells

231 Soluble heparinase-sensitive GAGs from Idua(-/-) BM and specific

232 tment of chondrocytes with either heparin or heparinase significantly reduced attachment to type XI c

233 eparin, heparan sulfate, sodium chlorate and heparinase, the low-density lipoprotein (LDL) receptor-r

234 L1 to diminish its intravascular function or heparinase to release CXCL1 from endothelial proteoglyca

235 and activated partial thromboplastin time in heparinase-treated samples in vitro.

236 In heparinase-treated samples, coagulation factor XII activ

237 spurious correlations between miRNAs in non-heparinase-treated samples.

238 cetic anhydride-d6 is followed by exhaustive heparinase treatment and disaccharide analysis by hydrop

239 a cellular uptake was blocked by heparin and heparinase treatment but not by LRP1 deficiency.

240 of 125I-bFGF to K562 cells was sensitive to heparinase treatment but not to chondroitinase treatment

241 and nonrheumatoid synovia was determined by heparinase treatment followed by an in situ binding assa

242 on assay demonstrated that entry blockage by heparinase treatment included the membrane fusion step.

243 Heparinase treatment of serial plasma and serum samples

244 Heparinase treatment of the cells did not abrogate this

245 cellular split-luciferase assay showed that heparinase treatment or adding heparin in culture medium

246 After heparinase treatment, LPL binding to apoB17-DAF cells wa

247 In cell surface binding assays, heparinase treatment, which degrades HS, or the knockout

248 Heparinase treatment, which digests anionic sulfated gly

249 heparan sulfate was depressed by chlorate or heparinase treatment.

250 ased 2-10-fold when the HSPG were removed by heparinase treatment.

251 Furthermore, this effect was blocked by heparinase treatment.

252 were still able to produce abundant NO after heparinase treatment.

253 ce heparin-like glycosaminoglycans using the heparinases, we recently have elaborated a mass spectrom

254 Finally, heparinases were used to identify and quantify GlcNH(3)(

255 e with heparin or pretreatment of cells with heparinase, which destroys heparan sulfate.

256 in (LRP), but were almost totally blocked by heparinase, which removes the sulfated glycosaminoglycan

257 Intravenous pretreatment of mice with heparinases, which specifically cleave heparan sulfate m

258 lowing the kinetic of heparin degradation by heparinase with a good correlation with results obtained