ライフサイエンスコーパス: barnase

1 Nase Sa, 44 in RNase T1, and 58 in RNase Ba (barnase).

2 fragment 6-85, chymotrypsin inhibitor 2, and barnase).

3 in our earlier study on charge mutations in barnase).

4 of five residues of barstar known to contact barnase.

5 al importance to the structural integrity of barnase.

6 ved in the series of N-terminal fragments of barnase.

7 t discernibly alter the folding mechanism of barnase.

8 ive-state HX data and the folding pathway of barnase.

9 sin inhibitor, chymotrypsin inhibitor 2, and barnase.

10 1.0x10(-7) M and the binding was competed by barnase.

11 e inhibitor barstar that binds to the enzyme barnase.

12 tudies of 1H/2H-exchange of amide protons of barnase.

13 ith a higher rate constant than singly bound barnase.

14 -fold in the proenzyme relative to wild-type barnase.

15 egions involved in the binding of barstar to barnase.

16 racellular inhibitor of the endoribonuclease barnase.

17 her cell types from the cytotoxic effects of barnase.

18 single amino acid dramatically destabilizes barnase.

19 might be important for the interaction with barnase.

20 chaperones bind to a fully unfolded state of barnase.

21 n interaction energy caused by a mutation in barnase.

22 problems in defining the folding pathway of barnase.

23 of ubiquitin inserted into a surface loop of barnase.

24 acid identity with the well-studied protein barnase.

25 variant of the 110-amino acid ribonuclease, barnase.

26 rall structure was similar to that of intact barnase.

27 imental delta G values is 0.09 kcal/mol, for barnase 0.14 kcal/mol, for the synthetic coiled-coil 0.1

28 in vivo for two unrelated monomeric enzymes: barnase (a bacterial RNase) and TEM-1 beta-lactamase.

29 etroviral gag protein onto the C-terminus of barnase, a ribonuclease produced by Bacillus amiloliquif

30 ermodynamic data are available: T4 lysozyme, barnase, a synthetic leucine zipper, and a synthetic pep

31 ed can complement each other to reconstitute barnase activity.

32 For gammaII-crystallin, myoglobin, barnase, alpha-lactalbumin, and cytochrome c the foldons

33 g in aqueous solution at 298 K for wild-type barnase and 66 mutants.

34 Barnase and barstar are trivial names of the extracellul

35 a side-chain involved in the interaction of barnase and barstar are, however, always unfavourable an

36 functional stomium region and that chimeric barnase and barstar genes containing promoters that are

37 failure to form a complex between the mutant barnase and barstar has a lethal effect on host cells un

38 tic interactions to the binding stability of barnase and barstar were studied by the Poisson-Boltzman

39 o be dominated by electrostatics, not unlike barnase and barstar, another well-characterized protein-

40 tion of crowding on the binding stability of barnase and barstar, based on atomistic modeling of the

41 ically assisted binding in complexes such as barnase and barstar.

42 revisited the pulse labeling experiment with barnase and detected no stable folding intermediate.

43 on constant of the complex between denatured barnase and either chaperone is 5 x 10(-8) M.

44 agree very well with experimental results on barnase and four other proteins.

45 folding and unfolding kinetics of wild-type barnase and four representative mutants under a wide ran

46 The rate of association of barnase and GroEL was found to be highly dependent on io

47 nitial, transient, ionic interaction between barnase and GroEL, before hydrophobic binding occurs, al

48 stest phase is related to the association of barnase and GroEL.

49 rvation that the unfolding rate constants of barnase and its mutants were divergent or convergent as

50 of the interaction between the ribonuclease barnase and its natural polypeptide inhibitor barstar.

51 he association of the bacterial ribonuclease barnase and its polypeptide inhibitor barstar which shed

52 Anthers containing the TA56/barnase and lectin/barstar genes also developed normally

53 At high ratios of GroEL to barnase and low ionic strength (less than 200 mM) this f

54 in understanding the early folding events of barnase and other proteins in general.

55 ion process from our previous simulations of barnase and protein A as well as new simulations of four

56 hat relies on the expression of a phytotoxic barnase and provides for male sterility.

57 agreement with those from similar studies of barnase and T4 lysozyme.

58 Anthers containing the TA56/barnase and TA20/barstar genes failed to dehisce as well

59 olvent model are validated using the Barstar-Barnase and the lysozyme-antibody D1.2 complexes, for wh

60 on the nature of the early folding state in barnase and therefore should have important implications

61 cence occurred in plants containing the TA56/barnase and TP12/barstar genes, indicating that barstar

62 previously fused two single-domain proteins, barnase and ubiquitin, such that the free energy stored

63 A small enzyme (barnase) and a ligand-binding polypeptide (GCN4) are fus

64 ribonuclease family, RNase T1 and RNase Ba (barnase), and with a member of the mammalian ribonucleas

65 r mouse dihydrofolate reductase or bacterial barnase, and found that unfolding of a precursor at the

66 ved in a range of non-crosslinked mutants of barnase as part of a general enthalpy-entropy compensati

67 model for the major folding intermediate of barnase, as well as the detailed pathway from the native

68 ative-state hydrogen exchange experiments on barnase at pD 5.0 and 25 degrees C and identified a part

69 The largest fragment of barnase, B105, has interactions that resemble its foldin

70 rying length fused to the small ribonuclease barnase (Ba).

71 serine and cysteine proteases as well as the barnase-barstar and Rap1a-raf complexes.

72 ously including HIs in BD simulations of the barnase-barstar association reaction.

73 The results presented here suggest that the barnase-barstar binding sites are correctly aligned duri

74 allographic solution of the structure of the barnase-barstar complex and the development of methods f

75 We have crystallised three mutants of the barnase-barstar complex in which interactions across the

76 pecific residues and interactions within the barnase-barstar interface to the enthalpy of binding has

77 d energy changes for double mutations in the barnase-barstar system is fully accounted for by conside

78 s and for all of the double mutations in the barnase-barstar system.

79 due mutations in the D1.3-HEL, D1.3-E5.2 and barnase-barstar systems and for all of the double mutati

80 in one case the net effect is close to zero (barnase-barstar) and in one case electrostatics provides

81 e binding of four protein-protein complexes; barnase-barstar, human growth hormone and its receptor,

82 protected from ablation by the formation of barnase/barstar complexes.

83 In the first group, represented by barnase/barstar, electrostatics exerts strong orientatio

84 comparison with the crystal structure of the barnase:barstar C40/82A complex revealed subtle differen

85 studies and in the crystal structure of the barnase:barstar C40/82A complex.

86 We also devised a barnase-based conditional suicide switch to further lowe

87 e have simulated the thermal denaturation of barnase beginning from the average NMR structure.

88 Up to 4 mol of denatured barnase bind to 1 mol of tetrameric SecB.

89 iquitin (Ub) into the bacterial ribonuclease barnase (Bn), using peptide linkers from zero to 10 amin

90 is inserted into one of six surface loops of barnase (Bn).

91 st complex bnHis102-->Ala-bsTyr29-->Phe (bn, barnase; bs, barstar), deletes a van der Waals packing i

92 cterised a series of C-terminal fragments of barnase by different biophysical techniques to find out

93 on in binding caused by the R59E mutation in barnase can be partly reversed by changing Glu-76 of bar

94 Using a transient system, we first show that barnase can be split into two inactive peptide fragments

95 nstrate that the extracellular ribonuclease, barnase, can be engineered into two complementary fragme

96 ion because their substrates are very small: barnase, carbonic anhydrase, glutathione S-transferase,

97 mutational data of the 15 proteins: barstar, barnase, chymotrypsin inhibitor 2 (CI2), Src SH3 domain,

98 The barnase coding information is divided and distributed at

99 promoter, PrMC2, was used to drive modified barnase coding sequences (barnaseH102E, barnaseK27A, and

100 stant of > 3 x 10(-4) M for the GroEL-native barnase complex.

101 ed with the anti-lysozyme antibody E5.2, and barnase complexed with barstar.

102 The four barnase complexes have native-like structure as shown by

103 thermodynamic and kinetic properties of four barnase complexes, with the cleavage site at different p

104 Barstar, an inhibitor of the enzyme barnase, contains two phenylalanine residues, three tryp

105 Seven fragments of the 110-residue protein barnase, corresponding to the progressive elongation fro

106 with a gene encoding a lethal ribonuclease, barnase, demonstrating that the INPACT system provides e

107 of formation of the folding intermediate of barnase directly, but have analysed its reactivity and t

108 each parent protein (catalytic efficiency of barnase, DNA binding affinity and sequence specificity o

109 ingly, mutations throughout the structure of barnase do not significantly affect the folding rate, su

110 In the absence of ligand, the barnase domain is more stable and is therefore folded an

111 CN4, forcibly unfolding and inactivating the barnase domain.

112 We find that SecB interacts with barnase during its folding in a similar manner to its in

113 approach utilizes a plasmid system in which barnase expression is tightly controlled to keep the mut

114 obacco line was transformed, indicating that barnase expression was responsible for the reduced frequ

115 op (Pro27-Glu32) towards the binding site of barnase facilitate the formation of interface hydrogen b

116 tive-state hydrogen exchange experiment with barnase failed to detect any partially unfolded intermed

117 ray crystal structure of pBn reveals how the barnase fold is able to adapt to permutation, partially

118 This agrees with the proposed model for barnase folding, where the residual structure in small f

119 gs are consistent with the proposed model of barnase folding.

120 fetime of potential intermediate states upon barnase folding/unfolding in the submillisecond timescal

121 Conversely, barnase folds from a largely structured denatured state

122 Previous studies led to the conclusion that barnase folds through a very stable submillisecond inter

123 other results in the literature suggest that barnase folds through partially unfolded intermediates t

124 loci is achieved through coexpression of the barnase fragments and intein-mediated ligation of the ba

125 arents constitutively expressing each of the barnase fragments, then assaying their progeny for the p

126 A description of the folding pathway of barnase from the denatured to the native state can be co

127 Binding of cobalt to the gag zinc finger-barnase fusion protein introduced sufficient anisotropic

128 Barnase-GCN4 is thus a "natively unfolded" protein that

129 Barnase-GCN4 thus defines a modular approach for assembl

130 We introduced a cytotoxic TA56/barnase gene into tobacco plants together with three dif

131 ion is tightly controlled to keep the mutant barnase gene silent.

132 three progeny inheriting only the N-terminal barnase gene were male fertile.

133 The lethal gene used here is a CaMV 35S-barnase gene with an intron in the coding sequence (barn

134 types from the cytotoxic effects of the TA56/barnase gene.

135 When expression of the partial barnase genes was instead targeted to the tapetum, male

136 eir progeny for the presence of both partial barnase genes.

137 ighest information content for inhibition of barnase (H102K) has the substitution Y30W.

138 The unfolding and folding of protein barnase has been extensively investigated in bulk condit

139 The folding/unfolding pathway of barnase has been studied extensively using the protein e

140 Molecular dynamics simulations of barnase have been conducted both in water and in 8 M ure

141 the stability of the folding intermediate of barnase (I) in 2H2O under a variety of conditions and ca

142 burst-phase (submillisecond) intermediate of barnase, if it exists, can be only marginally more stabl

143 ase Sa and RNase St differ considerably from barnase in both sequence and structure, yet both show si

144 o give the correct heat capacity of unfolded barnase in solution, it is possible to approximate the e

145 A series of studies on the small protein barnase in the 1990s established it as a paradigm for pr

146 the refolding of a singly bound molecule of barnase in the complex with GroEL.

147 Line broadening in the NMR spectra of barnase in the presence of chaperone indicates binding o

148 sible lowering of the melting temperature of barnase in the presence of chaperone.

149 on GroEL, and on the thermal denaturation of barnase in the presence of GroEL and SecB.

150 The refolding of barnase in the presence of GroEL is multiphasic, the slo

151 nd 13.5 +/- 2.5 pN catalyze the unfolding of barnase in those experiments.

152 emperature molecular dynamics simulations of barnase in water.

153 The incorporation of a barnase-INT gene outside the left border appears to prov

154 r or a control vector from which most of the barnase-INT gene was deleted.

155 gene with an intron in the coding sequence (barnase-INT); the screenable marker is a pMAS-luciferase

156 nology to convert the cytotoxic ribonuclease barnase into an artificial zymogen that is activated by

157 in four stabilised mutants than in wild-type barnase, irrespective of the presence of a disulfide cro

158 Barnase is a multi-modular protein that folds via an int

159 In contrast, unfolding of native barnase is catalysed by > 1000-fold.

160 nce that proves that the folding kinetics of barnase is inconsistent with the absence of a folding in

161 Multiply bound barnase is less tightly bound and refolds with a higher

162 igation of the nature of the intermediate of barnase is needed.

163 Barnase is one of the few protein models that has been s

164 l conditions, as the folding intermediate of barnase is the most populated state in the complex.

165 or exchange of buried amide protons of bound barnase is the unfolding of the folding intermediate, wh

166 When expression of barnase is turned on, failure to form a complex between

167 The folding intermediate of barnase is, thus, a relatively discrete and compact enti

168 cs of the ribonuclease binase, homologous to barnase, is investigated with (15)N, (13)C NMR relaxatio

169 es that the Streptomyces enzymes do have the barnase-like irregular beta-bulge, making this an import

170 than the distance between the termini of the barnase loop.

171 s obtained from the RCSB database to bind to barnase, lysozyme, and trypsin using a previously derive

172 rength (greater than 600 mM) the majority of barnase molecules escaped binding and refolded free in s

173 st cells against the RNAse activity of those barnase mutants not properly inhibited by wild-type bars

174 n enthalpies for complexes between different barnase mutants with amino acid substitutions of the gen

175 barstar suppressors has been identified for barnase mutants with substitutions in two amino acid pos

176 e N- and C-termini of the small ribonuclease barnase (normally 27.2 A distant) with a single Cys resi

177 effect of ionic strength on the refolding of barnase on GroEL, and on the thermal denaturation of bar

178 ics shows that tissue-specific expression of barnase or the antisense RTS genes interrupts tapetal de

179 eries of mutants that destabilize either the barnase or ubiquitin domains.

180 acillus amyloliquefaciens ribonuclease gene, barnase, or the antisense of the RTS gene, is able to dr

181 both show significant sequence similarity to barnase over a region beginning at Gly53.

182 cells along with genes encoding both partial barnase peptides, a substantial reduction in luciferase

183 oves the paramagnetic-induced orientation of barnase, permitting the measurement of only (1)J(HN) sca

184 ragments and intein-mediated ligation of the barnase protein fragments.

185 f this structural element to the function of barnase raises the question of whether it may be present

186 n inhibitor (BPTI) and in the I76A mutant of barnase, represent very different environments for the w

187 Both chaperones bind the denatured state of barnase, so lowering the T(m) value.

188 nd stability of three protein model systems: barnase, spectrin, and T4 lysozyme.

189 observations for hen egg white lysozyme and barnase, suggest that EX2 kinetics should not be assumed

190 e have analysed the conformational states of barnase that are bound by the molecular chaperones GroEL

191 dynamics of denaturation of three mutants of barnase that contain cystine and the corresponding singl

192 minimal reaction pathway for the folding of barnase that involves two detectable folding intermediat

193 ogen-deuterium exchange of amide proteins of barnase that require global unfolding for exchange to oc

194 In unfolded barnase, the residual interactions lead to downward pK(a

195 siderably greater than observed for Trp71 in barnase, the Trp on which Y52W is based.

196 proximate the experimental thermodynamics of barnase thermal denaturation: melting temperature, width

197 ochondrial import experiments on the protein barnase, these results imply that forces between 11 +/-

198 one indicates binding of the native state of barnase to both GroEL and SecB, with a dissociation cons

199 fect of Gly mutations on the folding rate of barnase to investigate the secondary structure formation

200 Subsequent collapse of unfolded barnase to the exchange-protected folding intermediate w

201 e protein drives unfolding of the other in a barnase-ubiquitin fusion protein.

202 Both chaperones bound to native barnase under physiological conditions and catalyzed exc

203 te the folding/unfolding reaction of protein barnase under the action of mechanical force at the sing

204 We present a computational study of barnase unfolding during import into mitochondria throug

205 nal impairment of the bacterial ribonuclease barnase upon substitution of Gly52 or Gly53.

206 effective barrier analysis show that protein barnase verifies the Leffler-Hammond postulate under app

207 thm of the unfolding rate constant of native barnase vs. denaturant concentrations is not linear.

208 Cytotoxicity of the reconstituted barnase was demonstrated by crossing together parents co

209 of Bacillus amyloliquefaciens ribonuclease (barnase) was analyzed by two-dimensional nuclear magneti

210 The refolding of barnase when bound to SecB is strongly retarded but neve

211 lding transition state of the larger protein barnase, which folds by a multi-state mechanism, with th

212 distribution of Phi-values resembled that of barnase, which folds via an intermediate, rather than th

213 by molecular dynamics the denatured state of barnase, which has been studied by NMR spectroscopy.

214 a similar way to that of the D93N mutant of barnase, which lacks the D93-R69 salt-bridge present in

215 Such exchange required complete unfolding of barnase, which occurred in the complex with the chaperon

216 On mixing acid-denatured barnase with SecB in a stopped-flow spectrofluorimeter u