ライフサイエンスコーパス: TnC

1 TnC binding to I1-64 broadens NMR signals from the side

2 TnC was then injected over the biosensor surface for qua

3 TnC-extracted cardiac skinned fibers were reconstituted

4 es cardiac contraction in response to Ca(2+) TnC binding Ca(2+) initiates a cascade of protein-protei

5 A TnC mutant that exhibited increased Ca(2+) sensitivity c

6 ardiac troponin (nTn), troponin containing a TnC mutant, CBMII, in which the sole regulatory site in

7 In contrast, a TnC mutant with decreased Ca(2+) sensitivity caused by a

8 These results provide the first example of a TnC domain in which the presence of only one calcium ion

9 librium from a TnI-actin-Tm interaction to a TnC-TnI interaction that likely enhances calcium binding

10 operties for the interacting TnI helix 1 and TnC COOH-domain.

11 and the distance between TnI residue 117 and TnC residue 98 was measured with the following results:

12 al to 130nM and 56nM for TnC-TnI(96-131) and TnC-TnI(96-139), respectively.

13 ither the distance between TnI residue 6 and TnC residue 89 nor the photo-cross-linking site in TnC,

14 1-29 (I1-29), and residues 1-64 (I1-64), and TnC.

15 ved for the interacting TnI switch helix and TnC NH(2)-domain, contrasting with stable, highly protec

16 bic interactions between this TnI region and TnC's N-domain cleft.

17 C-terminal to the phosphorylation sites and TnC either directly, due to electrostatic repulsion, or

18 pproximately 12 kcal/mol), and wild-type apo TnC ( approximately 20 kcal/mol).

19 that the nature of the interactions between TnC and the TnI-TnT component differs significantly betw

20 uctural information on the interface between TnC and this region of TnI has been long in dispute.

21 movement of TnI's inhibitory region between TnC and actin.

22 h the following results: for both the binary TnC-TnI complex and the ternary troponin complex, this d

23 emphasize the difference between the binary TnC.TnI and the ternary troponin complexes and the impor

24 Upon Ca(2+) binding, TnC changes conformation and exposes a hydrophobic patch

25 Cross-linking to both TnC and TnT is reduced by prior phosphorylation of the T

26 he extension lies in close proximity to both TnC and troponin T (TnT).

27 -bound TnC (3.2 kcal/mol), E40A Ca(2+)-bound TnC ( approximately 12 kcal/mol), and wild-type apo TnC

28 ng are calculated for wild-type Ca(2+)-bound TnC ( approximately 8 kcal/mol), V44Q Ca(2+)-bound TnC (

29 approximately 8 kcal/mol), V44Q Ca(2+)-bound TnC (3.2 kcal/mol), E40A Ca(2+)-bound TnC ( approximatel

30 Toll-like receptor 4 signaling activated by TnC, promoting an increased inflammatory response, hepat

31 tion that likely enhances calcium binding by TnC.

32 the detachment of TnI to calcium binding by TnC.

33 the temporospatial expression of tenascin-C (TnC) in balloon-injured rat and porcine arteries.

34 Tenascin-C (TnC), an extracellular matrix protein, is transiently ex

35 e of dissociation of Ca(2+) from troponin C (TnC) and decreased crossbridge detachment rate on the ti

36 ntributions of Ca(2+) binding to troponin C (TnC) and myosin binding to actin in activating thin fila

37 m-dependent interactions between troponin C (TnC) and other thin and thick filament proteins play a k

38 eraction between rabbit skeletal troponin C (TnC) and rabbit skeletal troponin I (TnI) regulatory reg

39 e-based sensor, carrying Opsanus troponin C (TnC) as the Ca(2+)-binding moiety, has two binding sites

40 Troponin C (TnC) belongs to the superfamily of EF-hand (helix-loop-h

41 ng of residues 1-64 (I1-64) with troponin C (TnC) by isothermal titration microcalorimetry and cross-

42 ivity, and Ca(2+) binding to the troponin C (TnC) component reverses the inhibition.

43 eleost fish, have two paralogous troponin C (TnC) genes that are expressed in the heart differentiall

44 ent and Ca(2+) dissociation from troponin C (TnC) have been hypothesized to rate-limit myofilament in

45 tation of the C-terminal lobe of troponin C (TnC) in skeletal muscle cells as a step toward elucidati

46 TnI) is thought to interact with troponin C (TnC) in the presence of Ca(2+) and with actin in the abs

47 ss converting calcium binding by troponin C (TnC) into interactions between thin and thick filaments.

48 Troponin C (TnC) is an EF-hand Ca(2+) binding protein that regulates

49 Cardiac troponin C (TnC) is composed of two globular domains connected by a

50 Troponin C (TnC) is implicated in the initiation of myocyte contract

51 s reconstituted with cardiac TnI.troponin C (TnC) or ssTnI.TnC significantly increased Ca(2+) sensiti

52 for cardiac muscle contraction, troponin C (TnC) stands out as an obvious and versatile target to re

53 fect of sarcomere length (SL) on troponin C (TnC) structure during Ca2+ activation in single glycerin

54 Calcium binding to the troponin C (TnC) subunit causes a change in its dynamics that leads

55 n enhances Ca(2+) binding to the troponin C (TnC) subunit of the troponin complex.

56 ing in the NH(2)-lobe of subunit troponin C (TnC) was abolished by mutagenesis, and effects on cardia

57 The native troponin C (TnC) was extracted and replaced with either cardiac TnC

58 Native troponin C (TnC) was extracted from single, de-membranated rabbit ps

59 ched along four alpha helices of troponin C (TnC) was measured in permeabilized skeletal muscle fiber

60 e switch is a dimeric complex of troponin C (TnC), an allosteric sensor for Ca(2+), and troponin I (T

61 control of different isoforms of troponin C (TnC), F1 and F2, which are responsible for stretch and c

62 Troponin C (TnC), present in all striated muscle, is the Ca(2+)-acti

63 l is transmitted between TnI and troponin C (TnC), resulting in accelerated Ca(2+) release, remains u

64 plex consists of three subunits, troponin C (TnC), troponin I (TnI), and troponin T (TnT).

65 t skeletal troponin I (TnI) with troponin C (TnC), troponin T (TnT), tropomyosin (Tm) and actin were

66 Mutations in the Ca(2+) sensor, troponin C (TnC), were generated to increase/decrease the Ca(2+) sen

67 ized fluorescence from probes on troponin C (TnC).

68 e reflect changes in Ca bound to troponin C (TnC).

69 he regulatory domain of skeletal troponin C (TnC).

70 troponin-I (TnI) and residues in troponin-C (TnC) and in actin.

71 ssess Ca(2+) affinity in isolated cardiac (c)TnCs containing F27W and did not necessarily mirror the

72 light muscle has two isoforms of troponin C, TnC-F1 and F2, which are unusual in having only a single

73 to TnC with competition between actin and Ca-TnC for the binding of TnI.

74 In the presence of calcium TnC-F2 is very similar to the control with approximately

75 s extracted and replaced with either cardiac TnC (cTnC) or a mixture of cTnC and an inactive mutant c

76 substituting native TnC with either cardiac TnC (cTnC), a site I-inactive skeletal TnC mutant (xsTnC

77 A genetically engineered cardiac TnC mutant labeled at Cys-84 with tetramethylrhodamine-5

78 by double-cysteine mutants of human cardiac TnC with bifunctional rhodamine attached along either th

79 in which the sole regulatory site in cardiac TnC (site II) is inactivated (CBMII-Tn), or troponin con

80 xture of cTnC and an inactive mutant cardiac TnC (CBMII TnC).

81 mutations influence the affinity of cardiac TnC for cTnI (KC-I) or contractile kinetics during beta-

82 t degrees of compactness between the cardiac TnC and skeletal TnC in their respective ternary complex

83 Mutations of the cardiac TnC N-terminal regulatory domain have been shown to alte

84 Ca2+ bound cooperatively to cardiac TnC in conventional thin filaments but noncooperatively

85 in filaments but noncooperatively to cardiac TnC in minifilaments in the absence of myosin.

86 reduced by replacing native TnC with cardiac TnC (cTnC) or a site I-inactive skeletal TnC mutant (xsT

87 akening the interaction of cTnI with cardiac TnC.

88 p between the cTnC fibres and the cTnC:CBMII TnC fibres would be apparent.

89 ships when comparing the cTnC and cTnC:CBMII TnC fibres.

90 Ca2+] and compared with different cTnC:CBMII TnC ratios at a saturating [Ca2+].

91 nC and an inactive mutant cardiac TnC (CBMII TnC).

92 with elevated hepatic stellate cell-derived TnC and Toll-like receptor 4 expression was observed in

93 replaced native cTnC with a mutant cTnC (DM-TnC) that is incapable of binding Ca(2+).

94 c decrease in the maximal force (F(max)), DM-TnC incorporation induced much larger increases in EC(50

95 ogressive replacement of native cTnC with DM-TnC caused an expected monotonic decrease in the maximal

96 Further, to determine whether elevated TnC expression correlated with obesity-associated HCC, w

97 Endogenous TnC was extracted, and the preparations were reconstitut

98 When Tn contained engineered TnC mutants with weakened regulatory TnI interactions, t

99 molecular dynamics simulations of engineered TnC variants that increase or decrease Ca(2)(+) sensitiv

100 Rationally engineered TnC constructs corrected the abnormal Ca(2+) sensitiviti

101 n be corrected through rationally engineered TnC constructs, three thin filament protein modification

102 We demonstrate engineering TnC can specifically and precisely modulate cardiac cont

103 induced obesity HCC mouse model and examined TnC expression and liver injury.

104 In vitro, myofibroblasts did not express TnC mRNA under basal conditions.

105 ed the solution structure of the isolated F1 TnC C-terminal domain in the absence of calcium and have

106 calcium loaded, the C-terminal domain of F1 TnC is in an open conformation which allows binding to t

107 We have previously shown that F1 TnC is a typical collapsed dumbbell EF-hand protein that

108 f calcium the three actin filaments (TnC-F1, TnC-F2 and mammalian control) are almost indistinguishab

109 bsence of calcium the three actin filaments (TnC-F1, TnC-F2 and mammalian control) are almost indisti

110 In this study we used a fluorescent TnC to measure both the rate of Ca(2+) dissociation from

111 A fluorescent TnC was utilized to measure Ca(2+) binding to TnC in the

112 exchange with the N-domain of a fluorescent TnC(F29W).

113 ysis: K(d) being equal to 130nM and 56nM for TnC-TnI(96-131) and TnC-TnI(96-139), respectively.

114 elded equilibrium dissociation constants for TnC (plus Ca(2+)) binding to the C-terminal TnI regulato

115 he mid point pCa for the switch differed for TnC-F1 and F2 (pCa 6.9 and 6.0, respectively) consistent

116 This higher occupancy of the M-state for TnC-F1, which occurs above pCa 6.9, is consistent with t

117 y 89-113 and approximately 130-150 away from TnC and towards actin.

118 re both the rate of Ca(2+) dissociation from TnC and the rate of cross-bridge detachment from several

119 ees C), the rate of Ca(2+) dissociation from TnC was faster than the rate of cross-bridge detachment

120 However, Ca(2+) dissociation from TnC was not as temperature-sensitive as cross-bridge det

121 Unlike Ca(2+) dissociation from TnC, cross-bridge detachment varied in myofibrils from d

122 man cardiac troponin I and troponin C (HCTnI.TnC) complex showed a decrease in the Ca(2+) sensitivity

123 idues play a crucial role in determining how TnC responds to calcium.

124 The distances to residues 12 and 41 in TnC's N-domain are both considerably longer than those p

125 Changes in TnC may also correlate with cardiac and various other mu

126 concentrations and conformational changes in TnC measured using a fluorescent probe, which provide ev

127 tances from this TnI residue to cysteines in TnC mutants were measured and used to localize this resi

128 However, the calcium-dependent increase in TnC(F29W) fluorescence could be reversed by addition of

129 preceding the first calcium-binding loop in TnC(F29W), was unchanged by addition of magnesium, but i

130 Force-pCa relationships in TnC((E59D,D75Y)) reconstituted rabbit psoas fibers and f

131 sidue 89 nor the photo-cross-linking site in TnC, Ser133, changes with Ca(2+), in support of the noti

132 n contrast to crystal structures of isolated TnC, the D and E helices are not collinear.

133 ever, the mutations that sensitized isolated TnC to calcium did not necessarily increase the calcium

134 of these mutations that sensitized isolated TnC to calcium on (1) the calcium binding and exchange w

135 mutant TnIs cross-link to both the isolated TnC N-domain and whole TnC.

136 the fast skeletal troponin molecule and its TnC subunit in the calcium saturated and depleted states

137 re recorded in intact KI-TnC-A8V(+/-) and KI-TnC-A8V(+/+) cardiomyocytes.

138 d fibers increased with mutant gene dose: KI-TnC-A8V(+/+)>KI-TnC-A8V(+/-)>wild-type, whereas KI-TnC-A

139 ed with mutant gene dose: KI-TnC-A8V(+/+)>KI-TnC-A8V(+/-)>wild-type, whereas KI-TnC-A8V(+/+) relaxed

140 containing the A8V mutation (heterozygote=KI-TnC-A8V(+/-); homozygote=KI-TnC-A8V(+/+)) were character

141 (heterozygote=KI-TnC-A8V(+/-); homozygote=KI-TnC-A8V(+/+)) were characterized by echocardiography and

142 actile transients were recorded in intact KI-TnC-A8V(+/-) and KI-TnC-A8V(+/+) cardiomyocytes.

143 Three-month-old KI-TnC-A8V(+/+) mice displayed decreased ventricular dimens

144 troponin C ( approximately 21%) into the KI-TnC-A8V(+/-) cardiac myofilament.

145 V(+/+)>KI-TnC-A8V(+/-)>wild-type, whereas KI-TnC-A8V(+/+) relaxed more slowly on flash photolysis of

146 , and enhanced systolic function, whereas KI-TnC-A8V(+/-) mice displayed cardiac restriction at 14 mo

147 The fluorescently labeled TnC was sensitive to both Ca(2+) dissociation and cross-

148 In fibres, M82Q TnC further slowed relaxation in low [P(i)] conditions b

149 d Ca(2+) dissociation rate in solution (M82Q TnC) also increased the Ca(2+) sensitivity of steady-sta

150 ased on the structure and dynamics of mutant TnC provide rationale for binding trends observed in GOF

151 In Tn complexes containing TnI121 and mutant TnCs with a single cysteine at positions 12, 48, 82, 98,

152 Native TnC was replaced by double-cysteine mutants of human car

153 in filaments was reduced by replacing native TnC with cardiac TnC (cTnC) or a site I-inactive skeleta

154 n rabbit psoas fibres by substituting native TnC with either cardiac TnC (cTnC), a site I-inactive sk

155 Ca(2+) dissociation rate in solution (NHdel TnC) decreased the Ca(2+) sensitivity of steady-state fo

156 ions by approximately twofold, whereas NHdel TnC had no significant effect on relaxation.

157 nker plasticity is important, the ability of TnC to function in muscle contraction can be correlated

158 change) to elucidate the binding affinity of TnC with Ca(2+) and, more importantly, to determine the

159 Furthermore, computational analysis of TnC showed the Ca(2+)-binding pocket as an active region

160 Binding of TnC to bisphosphorylated I1-64 does not broaden these NM

161 ted within the N-domain hydrophobic cleft of TnC in the presence of Ca2+, and that it moves out of th

162 ures of skeletal muscle troponin composed of TnC (the sensor), TnI (the regulator), and TnT (the link

163 Tn is composed of TnC, TnI, and TnT.

164 hesis and to investigate the conformation of TnC in the intact troponin complex and in solution, we a

165 tended rather than a compact conformation of TnC.

166 5-IAEDANS) was covalently linked to Cys98 of TnC and free TnI peptides were added.

167 xtremely tightly to the C-terminal domain of TnC and weakly to the N-terminal domain.

168 Ca(2+) binding to the regulatory domain of TnC removes the inhibitory effect by TnI on the contract

169 sion interacts with the N-terminal domain of TnC stabilizing Ca(2+) binding and that phosphorylation

170 8), and Met(81)) in the regulatory domain of TnC were individually substituted with polar Gln, to exa

171 at anchors I1-64 to the C-terminal domain of TnC.

172 TnI interacts with the N-terminal domain of TnC.

173 is predominantly to the N-terminal domain of TnC.

174 tants cross-link to the N-terminal domain of TnC.

175 for the residues linking the two domains of TnC, as well as for the inhibitory peptide residues prec

176 ces NMR signals arising from both domains of TnC, providing evidence that the N-terminal extension of

177 oute for propagating signals from one end of TnC to the other.

178 nvestigated the temporospatial expression of TnC by myofibroblasts after vascular injury.

179 investigate the temporospatial expression of TnC in injured arteries.

180 Moreover, expression of TnC L48Q after the MI therapeutically enhances cardiac f

181 n (MI) model of heart failure, expression of TnC L48Q before the MI preserves cardiac function and pe

182 used to determine the in vitro expression of TnC.

183 Extraction of TnC-reduced I-band and overlap Ca in rigor fibers at pCa

184 at the second EF-hand, but not the first, of TnC is responsible for the competitive magnesium binding

185 air was introduced into the first EF-hand of TnC(F29W), the fluorescence of G34DTnC(F29W) increased u

186 it is localized near the B and C helices of TnC's N-terminal domain.

187 The E helix of TnC makes an angle of about 45 degrees with the thin fil

188 in the regulatory Ca(2+)-binding Site II of TnC, TnC((E59D,D75Y)).

189 e effects observed in the calcium loading of TnC.

190 In relaxed muscle, the N-terminal lobe of TnC is less closed than in crystal structures of the Ca(

191 by X-ray crystallography, that this lobe of TnC is part of a well-defined troponin domain called the

192 To determine the cellular mechanism of TnC signaling in promoting inflammation and hepatocyte e

193 terms of a new quantitative dynamic model of TnC-TnI allostery.

194 The relative spatial orientation of TnC domains bound to TnI was calculated from measured re

195 movement of the Met121 region of TnI out of TnC's N-domain cleft is essential for the occurrence of

196 , residues approximately 114-125 move out of TnC's N-terminal domain hydrophobic cleft and trigger th

197 thin the regulatory Ca(2+)-binding pocket of TnC that hinders the relay of the thin filament calcium

198 TnI switch helix, in a hydrophobic pocket of TnC upon activation by Ca ions.

199 The functional properties of TnC and its ability to bind Ca(2+) have significant regu

200 Thus, both the intrinsic properties of TnC and its interactions with the other contractile prot

201 hat control the Ca(2+) binding properties of TnC and other EF-hand proteins are not completely unders

202 However, the mechanistic role of TnC signaling in the development of HCC remains unknown.

203 s 1-192) increased the Ca(2+) sensitivity of TnC on the thin filament, whereas the dilated cardiomyop

204 ltaK210, decreased the Ca(2+) sensitivity of TnC on the thin filament.

205 gnesium decreased the calcium sensitivity of TnC(F29W) and G34DTnC(F29W) approximately 2- and 6-fold,

206 red how a C-terminal calcium binding site of TnC can activate the thin filament.

207 ybridization revealed that the major site of TnC expression early after vascular injury was the adven

208 soas fibers and fluorescence spectroscopy of TnC((E59D,D75Y)) labeled with 2-[(4'-iodoacetamide)-anil

209 due with respect to the crystal structure of TnC.

210 those predicted by the crystal structure of TnC.TnI(1-47), supporting an extended rather than a comp

211 e of actomyosin ATPase activity than that of TnC or the Tn complex.

212 + removal but remains within the vicinity of TnC.

213 ng cross-bridge binding has little effect on TnC structure at any SL or level of activation.

214 tand the structural basis of their impact on TnC function.

215 transient opening of a hydrophobic patch on TnC's surface, to which a helix of another subunit, trop

216 ments reported in the literature for partial TnC replacement, increased [P(i)], increased [ADP], and

217 h HCC showed significant increases in plasma TnC compared with healthy volunteers and patients with l

218 h obesity-associated HCC, we measured plasma TnC in obese patients with various levels of liver injur

219 shown that the extracellular matrix protein, TnC, regulates cell migration.

220 Expression of recombinant TnC((E59D,D75Y)) in isolated rat cardiomyocytes induced

221 48 and 82 on opposite sides of troponin-C's (TnC's) N-terminal regulatory hydrophobic cleft photo-cro

222 helix that attaches to the Ca(2+)-saturated TnC NH(2) domain.

223 he relative orientation of calcium-saturated TnC domains when bound to cTnI, (1)H-(15)N residual dipo

224 y, our smartly formulated Ca(2+)-sensitizing TnC (L48Q) enhances heart function without any adverse e

225 serve as a bridge between the Ca(2+) sensor (TnC) and the actin filament.

226 trated asymmetrically by the dumbbell-shaped TnC subunit.

227 actness between the cardiac TnC and skeletal TnC in their respective ternary complexes and the fact t

228 rdiac TnC (cTnC), a site I-inactive skeletal TnC mutant (xsTnC), or mixtures of native purified skele

229 iac TnC (cTnC) or a site I-inactive skeletal TnC mutant (xsTnC).

230 sTnC) and a site I- and II-inactive skeletal TnC mutant (xxsTnC).

231 nC), or mixtures of native purified skeletal TnC (sTnC) and a site I- and II-inactive skeletal TnC mu

232 aced by mixtures of purified rabbit skeletal TnC (sTnC) and recombinant rabbit sTnC (D27A, D63A), whi

233 Teleost-specific TnC paralogs have not yet been functionally characterize

234 d with cardiac TnI.troponin C (TnC) or ssTnI.TnC significantly increased Ca(2+) sensitivity of force

235 When complexed with cardiac TnI.TnC or ssTnI.TnC, both TnT1 DCM mutations strongly decreased maximal

236 tituted with either cardiac TnI.TnC or ssTnI.TnC, significantly decreased Ca(2+) sensitivity of force

237 binding site in the NH(2) domain of subunit TnC.

238 Overall, these studies suggest TnC/Toll-like receptor 4 signaling as an important regul

239 e development of inotropic drugs that target TnC.

240 olved in the formation of neointima and that TnC facilitates migration of these cells during adventit

241 munohistochemical staining demonstrated that TnC expression began in adventitial myofibroblasts three

242 vidual mutants in cardiomyocytes showed that TnC(D75Y) was able to recapitulate the TnC((E59D,D75Y))

243 s to actin, the TnT-TnI coiled-coil, and the TnC COOH domain that contains the regulatory Ca(2+) site

244 Both the IT coiled coil and the TnC E helix tilt by about 10 degrees on muscle activatio

245 clearer view of the interactions between the TnC and TnI proteins of the troponin complex.

246 nd kinetics of the opening transition of the TnC hydrophobic patch.

247 een the open and closed conformations of the TnC molecule, which provides an indirect mechanism for t

248 2+) binding and conformational change of the TnC molecule.

249 binding of calcium to sites I and II of the TnC subunit results in a set of structural changes in th

250 between the Ca(2+) binding properties of the TnC(F29W) mutants and the solvent accessibility of the h

251 e global N-terminal Ca(2+) affinities of the TnC(F29W) mutants varied approximately 2340-fold, while

252 that TnC(D75Y) was able to recapitulate the TnC((E59D,D75Y)) phenotype, whereas TnC(E59D) was functi

253 he regulatory Ca(2+)-binding Site II of TnC, TnC((E59D,D75Y)).

254 the component of the troponin complex, TnI, TnC, TnT, that is responsible for inhibition of actomyos

255 n TnT and other thin filament proteins, TnI, TnC and Tm.

256 minent with a hybrid troponin (skeletal TnI, TnC, and cardiac TnT) than with all cardiac troponin.

257 onin itself consists of three subunits, TnI, TnC, and TnT, widely characterized as being responsible

258 lobular head region with the regulatory TnI- TnC complex with a tail anchoring it within the thin fil

259 rate of Tn dissociation is by favoring a TnI-TnC interaction over a TnI-actin-Tm interaction.

260 anogen bromide digestion of the covalent TnI-TnC complex formed from intact troponin demonstrates tha

261 t model of the Ca(2+)-saturated skeletal TnI-TnC complex in which the inhibitory region is modeled as

262 ap region increased proportionately with TnI-TnC regulatory affinity.

263 When complexed with cardiac TnI.TnC or ssTnI.TnC, both TnT1 DCM mutations strongly decre

264 , when reconstituted with either cardiac TnI.TnC or ssTnI.TnC, significantly decreased Ca(2+) sensiti

265 were complexed with recombinant cardiac TnT/TnC and exchanged into skinned rat cardiac trabeculae.

266 The binding of Ca(2+) to TnC initiates a cascade of conformational changes involv

267 at interact with TnC: I1-18 does not bind to TnC whereas the C-terminal region of unphosphorylated I1

268 hing domain from actin after Ca2+ binding to TnC (the Ca2+ sensor of troponin that relieves inhibitio

269 by corresponding changes in Ca2+ binding to TnC and that strong cross-bridge binding has little effe

270 nC was utilized to measure Ca(2+) binding to TnC in the physiologically relevant biochemical model sy

271 I1-64 binding to TnC influences NMR signals arising from both domains of

272 and predicts cooperative calcium binding to TnC with competition between actin and Ca-TnC for the bi

273 released from actin upon calcium binding to TnC, while TnT performs a structural role forming a glob

274 d cycling cross-bridges on Ca(2+) binding to TnC.

275 role in modulating the binding of calcium to TnC in increasingly complex biochemical systems.

276 -maleimidobenzophenone photo-cross-linked to TnC but not to actin in both the presence and absence of

277 Ca(2+) enhances the cross-linking to TnC.

278 inds to actin in the absence of Ca(2+) or to TnC in the presence of Ca(2+).

279 (4) BP-TnI121 photocrosslinked to TnC with a small degree of Ca(2+)-dependence, and did no

280 All the labeled mutants photocrosslinked to TnC with varying degrees of Ca(2+)-dependence, and none

281 (5) BP-TnI155 and TnI179 photocrosslinked to TnC, TnT and actin, but all with low yields.

282 Photocrosslinking to TnC was reduced in the ternary versus the binary complex

283 s: (1) BP-TnI6 photocrosslinked primarily to TnC with a small degree of Ca(2+)-dependence in all the

284 in alignment tensor orientation for the two TnC domains supports restriction of domain motion in the

285 the reported calcium affinities for the two TnCs.

286 mulations corroborate that cardiac wild-type TnC does not open on timescales relevant to contraction

287 lloon-injured arteries, markedly upregulated TnC mRNA.

288 The engineered sensor, called NTnC, uses TnC as the Ca(2+)-binding moiety, inserted in the mNeonG

289 By one week, TnC expression reached the developing neointima.

290 late the TnC((E59D,D75Y)) phenotype, whereas TnC(E59D) was functionally benign.

291 ate and 10-15% in the fully on M-State while TnC-F1 has almost 50% in each of the C and M states.

292 to both the isolated TnC N-domain and whole TnC.

293 itivity approximately 1.7-fold compared with TnC(F29W) at 1 mm [magnesium](free) and approximately 3.

294 on (1) the calcium binding and exchange with TnC in increasingly complex biochemical systems and (2)

295 y affected calcium binding and exchange with TnC in increasingly complex biochemical systems, indicat

296 the N-terminal half of TnI to interact with TnC in a Ca(2+)-independent manner, while the C-terminal

297 N-terminal regions of TnI that interact with TnC: I1-18 does not bind to TnC whereas the C-terminal r

298 Ca(2+), reflecting closer interactions with TnC under these conditions.

299 and the preparations were reconstituted with TnC fluorescently labeled with 5'ATR.

300 away from actin and interacted strongly with TnC in the presence of Ca(2+).