ライフサイエンスコーパス: heparinase

1 to treatment of macrophages with trypsin or heparinase.

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

3 sensitive to both chondroitin ABC lyase and heparinase.

4 main, and by incubating the blastocysts with heparinase.

5 excess heparan sulfate and by treatment with heparinase.

6 sion was reversed by tetrahydrolipstatin and heparinase.

7 which was reversed by antifibrinolytics and heparinase.

8 y soluble heparin and by treating cells with heparinase.

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

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

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

12 broadest known substrate specificity 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 rin formation) greater than 1 minute between heparinase and standard thrombelastogram (TEG) is associ

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

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

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

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

26 Heparinases are bacterial enzymes that are powerful tool

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

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

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

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

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

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

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

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

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

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

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

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

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

40 The three heparinases derived from Flavobacterium heparinum are po

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

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

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

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

45 Heparinase digestion of the endothelial cell's glycocaly

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

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

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

49 end of oligosaccharides resulting from prior heparinase eliminative cleavage.

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

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

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

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

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

55 The heparinases from Flavobacterium heparinum are lyases tha

56 The heparinases from Flavobacterium heparinum are powerful t

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

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

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

60 Heparinase had no effect on adhesion to the peptide.

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

62 In addition, heparinases have significant therapeutic applications.

63 Heparinase/heparinitase treatment of the basolateral cel

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

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

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

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

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

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

70 Heparinase I activity is restored following hydroxylamin

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

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

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

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

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

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

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

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

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

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

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

82 60 nm) and that calcium is able to activate heparinase I catalytically.

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

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

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

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

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

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

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

90 Heparinase I from Flavobacterium heparinum has important

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

108 Heparinase I was immobilized onto agarose beads via cyan

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

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

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

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

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

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

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

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

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

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

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

120 n-like glycosaminoglycan depolymerization by heparinase I.

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

122 cofactor without altering heparin binding to heparinase I.

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

124 ve mechanism of depolymerization of HLGAG by heparinase I.

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

126 microenvironment for the catalytic domain of heparinase I.

127 zation of heparin-like glycosaminoglycans by heparinase I.

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

129 sidues together form the catalytic domain of heparinase I.

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

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

132 was abrogated by pretreatment of cells with heparinase I.

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

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

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

136 from Flavobacterium heparinum, specifically heparinases I and II.

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

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

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

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

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

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

143 Heparinase II activity is restored following hydroxylami

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

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

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

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

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

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

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

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

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

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

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

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

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

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

158 Heparinase II is unique among the heparinases in that it

159 Heparinase II is unique among the three enzymes because

160 However, heparinase II preincubation with heparan sulfate was una

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

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

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

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

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

166 ysteine 189 are functionally unimportant for heparinase II.

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

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

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

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

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

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

173 Heparinase III and human heparanase specifically cleaved

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

175 95 and histidine 510, as being essential for heparinase III enzymatic activity.

176 Heparinase III is the unique member of the heparinase fa

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

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

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

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

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

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

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

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

185 Heparinase III treatment of the genital tract significan

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

202 as completely inhibited by pretreatment with heparinase III.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

219 Antifibrinolytics and heparinase partially reverse the abnormal clotting patte

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

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

222 Heparinase-pretreatment of LRP-deficient fibroblasts or

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

224 Blocking FGF1 receptors with heparinase reduced this effect.

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

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

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

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

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

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

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

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

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

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

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

236 Heparinase treatment of the cells did not abrogate this

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

238 heparan sulfate was depressed by chlorate or heparinase treatment.

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

240 Furthermore, this effect was blocked by heparinase treatment.

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

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

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

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

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

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