ライフサイエンスコーパス: phosphoenzyme

1 higher affinity for the cation-free form of phosphoenzyme.

2 suggests that GFP-Spf1 accumulated the E1~P phosphoenzyme.

3 es at a slow rate involving alkali-resistant phosphoenzyme.

4 of phosphorylation and the maximal level of phosphoenzyme.

5 he global structure of this low fluorescence phosphoenzyme.

6 e 6-phosphate, followed by hydrolysis of the phosphoenzyme.

7 r polypeptide also formed a Ca(2+)-dependent phosphoenzyme.

8 binding to and release from the Na,K-ATPase phosphoenzyme; a value of 130 s(-1) for k(2), a rate con

9 ith an initial burst involving alkali labile phosphoenzyme (absent in D1027N and C983A/C985A mutants)

10 measured low-affinity ATP binding to stable phosphoenzyme analogues, demonstrating that the E1-P to

11 s so primarily by enhancing the level of the phosphoenzyme and only when ATP is used as the phosphate

12 es the first characterization of a P5 ATPase phosphoenzyme and points to Ca(2+) as a modifier of its

13 gher Na+ concentrations reduced the level of phosphoenzyme and stimulated Na-ATPase activity in the a

14 of 9 + 2 cilia and flagella via a network of phosphoenzymes and regulatory proteins.

15 However, comparable levels of phosphoenzyme are reached in the presence of both cation

16 LPCAT1 was identified as a phosphoenzyme as Ser(178) within a phosphodegron was ide

17 Chasing the phosphoenzyme at 0.4 M KCl and 2 degrees C with ADP reve

18 gamma-(32)P]ATP of a (32)P-labeled transient phosphoenzyme at the catalytic site of Drs2p.

19 pproximately 20 times faster than subsequent phosphoenzyme breakdown.

20 e enzyme while retaining the ability to form phosphoenzyme by utilization of P(i).

21 Alkali labile phosphoenzyme (catalytic intermediate of P-ATPases) acco

22 Contrary to SERCA, ATP7B phosphoenzyme cleavage shows much lower temperature depe

23 re on the characteristics of E(1), E(2), and phosphoenzyme conformations were examined by measuring b

24 ) selective inward facing sites, whereas the phosphoenzyme contains K(+) selective outward facing sit

25 Formation of this phosphoenzyme could be detected only if Cdc50p was co-ex

26 -phospholamban had any effect on the rate of phosphoenzyme decay relative to Ca-ATPase expressed alon

27 the E.Ca to E'.Ca transition or the rate of phosphoenzyme decay.

28 ormal structural transitions associated with phosphoenzyme decay.

29 The phosphoenzyme (E-P) binds PRPP with a KD of 0.6 microM,

30 abled the isolation and analysis of both the phosphoenzyme (E-P) formation and hydrolysis step.

31 bound nucleotide (E x ATP), and formation of phosphoenzyme (E-P).

32 eacts with 0.01-2.00 mM ATP to form covalent phosphoenzyme (E-P).

33 eady-state accumulation of the ADP-sensitive phosphoenzyme (E1P), and a rapid phase of EGTA-induced p

34 +),K(+)-ATPase, with highest affinity to the phosphoenzyme (E2P) forms.

35 yme (E1P), and a rapid phase of EGTA-induced phosphoenzyme (E2P) hydrolysis.

36 a corresponding decline in the steady-state phosphoenzyme (EP) level.

37 Chasing the phosphoenzyme (EP) with 1.66 mM ADP 10 ms after the star

38 Decay of [(32)P]phosphoenzyme following chase with non-radioactive ATP o

39 nt for Ca(2+) binding (distal allostery) and phosphoenzyme formation (direct activation).

40 bile and driven by Brownian motion to elicit phosphoenzyme formation and calcium transport, respectiv

41 t of Cys(119) by alanine or serine abrogates phosphoenzyme formation and phosphohydrolase activity.

42 f the ATPase mechanism (i.e., Ca2+ dependent phosphoenzyme formation and thapsigargin sensitivity) ar

43 Measurements of the time course of phosphoenzyme formation at 0 degrees C, using 1 microM M

44 affinity of ATP, (ii) the maximal extent of phosphoenzyme formation by ATP, (iii) the rate of steady

45 ispensable for phosphohydrolase activity and phosphoenzyme formation by BVP.

46 orm an intermediate state, which facilitates phosphoenzyme formation from ATP upon occupancy of the s

47 Steady-state phosphoenzyme formation from ATP was reduced in mutants

48 cation-dependent ATP hydrolytic activity and phosphoenzyme formation from ATP.

49 the enzyme is not the rate limiting step for phosphoenzyme formation from E.Na3.

50 ons of ADP are shown to increase the rate of phosphoenzyme formation of E. coli succinyl-coenzyme A (

51 amics of E, E x ATP, or E-P, indicating that phosphoenzyme formation or nucleotide binding result in

52 amban had any effect on the apparent rate of phosphoenzyme formation relative to that of Ca-ATPase ex

53 Although Gln-446 is not essential for the phosphoenzyme formation step, it plays an important role

54 hosphate (P(i)) release concomitant with the phosphoenzyme formation studies showed that L37A-phospho

55 ity and Ca(2+) transport by interfering with phosphoenzyme formation with ATP or P(i).

56 s are tightly constrained by ATP binding and phosphoenzyme formation, and this constraint must be ove

57 ver, the rates (but not the final levels) of phosphoenzyme formation, as well the rates of its hydrol

58 ot interfere with high-affinity ATP binding, phosphoenzyme formation, or phosphoenzyme hydrolysis.

59 From the copper dependence of phosphoenzyme formation, the mutants appear to have 2-3

60 at Ca(2+) efflux by LMCA1 is rate-limited by phosphoenzyme formation.

61 acid ensures productive partitioning toward phosphoenzyme formation.

62 tion of the GFP-Spf1 molecule that abolished phosphoenzyme formation.

63 ion and interference with Ca(2+) binding and phosphoenzyme formation.

64 in loss of copper resistance, transport and phosphoenzyme formation.

65 In crude membranes, the phosphoenzyme formed at steady state at 4 degrees C disp

66 etin increased the steady-state formation of phosphoenzyme from ATP or Pi, but higher quercetin decre

67 and the probability of calcium pump forming phosphoenzyme from bound P(i) (P(c) = 0.04 +/- 0.03) was

68 However, the formation of phosphoenzyme from E.Na3 with 1.0 mM ATP plus 2.0 mM ADP

69 present crystal structures representing the phosphoenzyme ground state (E2P) and a dephosphorylation

70 that at this resolution the low fluorescence phosphoenzyme had a structure similar to that of the nat

71 1.Ca to E1'.Ca transition and/or the rate of phosphoenzyme hydrolysis.

72 ity ATP binding, phosphoenzyme formation, or phosphoenzyme hydrolysis.

73 The level of ADP-sensitive phosphoenzyme (i.e. E1P-2Ca(2+)) observed in the transie

74 of ~0.3-1 mumol of P(i)/mg/min and formed a phosphoenzyme in a simple reaction medium containing no

75 rnover are accelerated by ATP binding to the phosphoenzyme in exchange for ADP.

76 dylserine, ATPase II would be accumulated as phosphoenzyme in the presence of ATP, resulting in the i

77 ke, the formation and decomposition of SERCA phosphoenzyme intermediate (E-P) in mouse cardiac homoge

78 ) ATPases utilize ATP through formation of a phosphoenzyme intermediate (E-P) whereby phosphorylation

79 a two-step mechanism that proceeds through a phosphoenzyme intermediate (E-P).

80 ng: 1) P5N-1 with bound Mg(II) at 2.25 A, 2) phosphoenzyme intermediate analog at 2.30 A, 3) product-

81 at despite sharing an HCxxxxxR(S/T) motif, a phosphoenzyme intermediate and a core alpha/beta-fold wi

82 ction of the cycle and also to form ATP from phosphoenzyme intermediate and ADP in the reverse direct

83 tion occurs in two steps: the formation of a phosphoenzyme intermediate and release of beta-D-fructos

84 n, activated MKP3 catalyzes formation of the phosphoenzyme intermediate approximately 100-fold faster

85 Our results demonstrate that ATP8A2 forms a phosphoenzyme intermediate at the conserved aspartate (A

86 The phosphoenzyme intermediate behaved in a chemically and k

87 rein is based on the formation of a covalent phosphoenzyme intermediate during substrate turnover.

88 sual among family members in that the common phosphoenzyme intermediate exists as a stable ground-sta

89 strates that Asp128 plays a role in both the phosphoenzyme intermediate formation (k2) and breakdown

90 s been proposed and herein investigated: (1) phosphoenzyme intermediate formation and (2) phosphoenzy

91 2+)- and Mn(2+)-dependent ATP hydrolysis and phosphoenzyme intermediate formation in forward (ATP) an

92 conclude that in addition to common (P-type) phosphoenzyme intermediate formation, SERCA and ATP7A/B

93 phosphoenzyme intermediate formation and (2) phosphoenzyme intermediate hydrolysis.

94 is explains why phosphoryl transfer from the phosphoenzyme intermediate in PTPases can only occur to

95 of a beta-PGM and the first view of the true phosphoenzyme intermediate in the HAD superfamily.

96 P hydrolysis indicated that formation of the phosphoenzyme intermediate is approximately 20 times fas

97 all of these NTPs, suggesting that a common phosphoenzyme intermediate is formed.

98 A phosphoenzyme intermediate may account for the dual func

99 Furthermore, studies with the phosphoenzyme intermediate of ATPase II and its recombin

100 ults indicate that the phosphate bond of the phosphoenzyme intermediate of H(+)-ATPases is labile in

101 the release of fructose-6-phosphate from the phosphoenzyme intermediate product complex.

102 ities were measured, and the turnover of the phosphoenzyme intermediate was close to the wild-type en

103 The pH-stability profile of the phosphoenzyme intermediate was consistent with phosphohi

104 The steady-state level of SERCA phosphoenzyme intermediate was increased 2.5-fold, and t

105 The phosphoenzyme intermediate was stable in 1 M NaOH and la

106 of N796 and E309 are both required to form a phosphoenzyme intermediate with ATP in the forward direc

107 ru-6-P + Pi) has been shown to proceed via a phosphoenzyme intermediate with His258 phosphorylated, a

108 Michaelis complex of an inactive mutant, the phosphoenzyme intermediate, and the product complex, a f

109 Mn(2+), and Zn(2+) stimulated formation of a phosphoenzyme intermediate, consistent with the transloc

110 cysteine side chain of the protein to form a phosphoenzyme intermediate, has been studied by combinin

111 cement general base in the hydrolysis of the phosphoenzyme intermediate, rescuing some of the activit

112 in proximity to the phosphoryl group of the phosphoenzyme intermediate, thus increasing the rate of

113 r defined by the isolation of a radiolabeled phosphoenzyme intermediate, which identified a conserved

114 he absence of copper, to form a low-turnover phosphoenzyme intermediate, with a conformation similar

115 atalytic site yielding stable analogs of the phosphoenzyme intermediate, with properties similar to E

116 nes through the formation and breakdown of a phosphoenzyme intermediate.

117 teine, and facilitating the formation of the phosphoenzyme intermediate.

118 ition state for the dephosphorylation of the phosphoenzyme intermediate.

119 transition states and directly observing the phosphoenzyme intermediate.

120 haracterized by formation and breakdown of a phosphoenzyme intermediate.

121 ere carried out to test the formation of the phosphoenzyme intermediate.

122 t phosphoryl transfer without formation of a phosphoenzyme intermediate.

123 nt engagement of the -phosphate yielding the phosphoenzyme intermediate.

124 uct complex after hydrolysis of the covalent phosphoenzyme intermediate.

125 of phosphotransferases that act via an acyl-phosphoenzyme intermediate.

126 participates in the dephosphorylation of the phosphoenzyme intermediate.

127 The structure is comparable to that of a phosphoenzyme intermediate: BeF(3)(-) is bound to Asp-11

128 Thereby, Ca(2+) and/or nucleotide-bound phosphoenzyme intermediates accumulate and undergo uncou

129 ther phosphatases that function via covalent phosphoenzyme intermediates, ComB can catalyze a transph

130 These results were confirmed in assays of phosphoenzyme intermediates.

131 k2 at serine 15 and the translocation of the phosphoenzyme into the cytosol.

132 trometry demonstrates that the alkali stable phosphoenzyme involves Ser(478) and Ser(481) (NMBD), Ser

133 of three sodium ions, and hydrolysis of the phosphoenzyme is associated with an influx of two potass

134 This phosphoenzyme is characterized by very low fluorescence

135 56A mutant enzyme were unsuccessful, but the phosphoenzyme is detected in the wild type, H390A, R255A

136 lity as well as the low fluorescence of this phosphoenzyme is due to a fluorescein-mediated cross-lin

137 37A-phospholamban decreased the steady-state phosphoenzyme level to a greater extent (45%) than did w

138 t affinity for Ca(2+) and an increase in the phosphoenzyme levels at steady state.

139 When steady-state Ca-ATPase phosphoenzyme levels were measured at 0 degrees C, using

140 e corresponded to 10% of total protein, with phosphoenzyme levels, catalytic turnover and Ca(2+) tran

141 om ATP or Pi, but higher quercetin decreased phosphoenzyme levels.

142 If formed. the phosphoenzyme-oxalate complex should be sufficiently sta

143 ed to produce a detectable level of the [32P]phosphoenzyme-oxalate complex.

144 other hand, subsequent (forward or reverse) phosphoenzyme processing is sensitive to activation ener

145 Previous work characterized an ultrastable phosphoenzyme produced first by labeling with fluorescei

146 results do not support the intermediacy of a phosphoenzyme-pyruvyl enolate complex in PEP mutase cata

147 nce that PEP mutase catalysis proceeds via a phosphoenzyme-pyruvyl enolate intermediate.

148 In contrast, the phosphoenzyme rapidly decayed to less than 20% when chas

149 ased by the addition of cold ATP, 90% of the phosphoenzyme remained stable after 5 s.

150 an inhibitory effect on the formation of the phosphoenzyme similar to that of Ca(2+) TheKmfor ATP in

151 2+)-free, Ca(2+)-bound, and actively cycling phosphoenzyme states (E2, E1, and EP).

152 Using the properties of the phosphoenzyme, the partial reaction steps of the transpo

153 However, the phosphoenzyme turnover (likely rate limited by the "Ca2.

154 The effects of Val-304 mutations on phosphoenzyme turnover are attributed to interference wi

155 x but dissociates with lower affinity as the phosphoenzyme undergoes a further conformational change

156 Here we demonstrate the formation of a BVP phosphoenzyme upon reaction with [gamma-(32)P]ATP and id

157 ed that the formation of the Spf1p catalytic phosphoenzyme was fast in a reaction medium containing A

158 tivity, suggesting that the formation of the phosphoenzyme was not the limiting step of the ATP hydro

159 2.7.4.1) of Escherichia coli, an N-P-linked phosphoenzyme was previously identified as the intermedi

160 To study the structural properties of this phosphoenzyme, we used cryoelectron microscopy of two-di

161 Incubation of the (32)P-labeled phosphoenzyme with 3-PGA resulted in the disappearance o

162 metals drive the formation of an acid-stable phosphoenzyme with apparent affinities similar to those