ライフサイエンスコーパス: ecotin

1 e reactive-site peptide bonds in eglin C and ecotin.

2 hydrogen bond with the P2 carbonyl group of ecotin.

3 ants (K(i)s) comparable to that of wild-type ecotin.

4 addition of a matriptase-specific variant of ecotin.

5 10(4) times stronger than that of wild-type ecotin.

6 is specific for proteins that interact with ecotin.

7 The ecotin 60X4 library, in which residues 67 to 70 of ecoti

8 The structure of the ecotin 67-70A, M84R mutant bound to trypsin shows large

9 Ecotin 67-70A, M84R which has four alanine substitutions

10 ptide was expressed at the amino terminus of ecotin, a dimeric macromolecular serine protease inhibit

11 Ecotin, a dimeric serine protease inhibitor from Escheri

12 607 in complex with a substitution mutant of ecotin, a panserine protease protein inhibitor secreted

13 Ecotin, a potent macromolecular inhibitor of serine prot

14 f complexes of trypsin N143H, E151H bound to ecotin A86H are determined at 2.0 A resolution via X-ray

15 , suggests that the protease binding loop of ecotin adopts a conformation mimicking that of the cleav

16 The inherent flexibility of ecotin allows it to accommodate these mutations and stil

17 ease inhibitor was realized by the use of an ecotin analogue containing allylglycine at position 84 i

18 Iodine treatment of ecotin analogue Met84(A)Gly resulted in the deactivation

19 ld be released from the protein complex with ecotin analogue Met84(A)Gly.

20 sizing system, permitting the elaboration of ecotin analogues containing allyglycine at the desired s

21 The derived ecotin analogues were capable of inhibiting bovine tryps

22 with the P1 residue of the bound inhibitors ecotin and bovine pancreatic trypsin inhibitor suggested

23 filled with eight consecutive amino acids of ecotin and demonstrates thrombin's preference for specif

24 nding site the molecular recognition between ecotin and its target serine proteases was probed.

25 The interaction between ecotin and target proteases with trypsin-like specificit

26 Ecotin and variant ecotins are subnanomolar inhibitors o

27 the conformation of several loop regions of ecotin are altered, resulting in the secondary site rele

28 Ecotin and variant ecotins are subnanomolar inhibitors of the MT-SP1 activa

29 is enzyme complexed to the protein inhibitor ecotin at 2.5 A resolution reveals a large primary bindi

30 d with the dimeric serine protease inhibitor ecotin at 2.5 A resolution reveals an extended cleft pro

31 trypsin at positions 143 and 151 and one on ecotin at position 86, anchor the metals and provide ext

32 e investigated with alanine substitutions in ecotin at Trp67, Gly68, Tyr69, Asp70, Arg108, Asn110, Ly

33 s than fivefold difference in the potency of ecotin binding to rat trypsin, suggesting that the exten

34 Ecotin binds to proteases with a chymotrypsin fold throu

35 Furthermore, GGH-ecotin can be cross-linked to a serine protease target,

36 Met84(A)Gly resulted in the deactivation of ecotin, caused by peptide backbone cleavage at its P1 re

37 rypsin resulted in enrichment as well as the ecotin consensus sequence WGFP at positions 67 to 70.

38 ulted in enrichment as well as the different ecotin consensus sequence WGYR at positions 67 to 70.

39 ecotin variant (mEcotin) and a single-chain ecotin dimer (scEcotin) were constructed to study the ef

40 that has an apparent molecular mass of a GGH-ecotin dimer with no observable protein degradation.

41 We used a variant of ecotin (ecotin-PKal), a macromolecular inhibitor of seri

42 -tetramer with three distinct interfaces: an ecotin-ecotin dimer interface, a larger primary ecotin-p

43 The variant ecotin encoded by this sequence inhibited rat trypsin at

44 The protease inhibitor ecotin fails to inhibit thrombin despite its broad speci

45 e synthesized by in vitro translation of the ecotin gene containing an engineered nonsense codon (TAG

46 Wild-type ecotin has an affinity of 1 nM for trypsin, while the op

47 efforts was taken to create potent bidentate ecotin inhibitors for trypsin and human urokinase-type p

48 ey amino acids with histidine at the trypsin-ecotin interface in the S2'/P2' pocket.

49 Ecotin is a dimeric serine protease inhibitor from Esche

50 Ecotin is a homodimeric protein from Escherichia coli th

51 In the presence of Ni(OAc)2 and MMPP, GGH-ecotin is cross-linked to give a species that has an app

52 restricted tetranomial diversity, we created ecotin libraries that are mutated at all 20 amino acid r

53 A comparison with two recently described ecotin-like genes from other bacteria suggests that thes

54 y percent of L. major-knockout lines for the ecotin-like serine peptidase inhibitor (ISP2; Deltaisp2/

55 Thrombin and ecotin M84R interact in two distinct surfaces.

56 ovine thrombin is determined in complex with ecotin M84R mutant at 2.5 A resolution.

57 A second generation library, ecotin M84R+60X4 including an additional methionine to a

58 Furthermore, ecotin M84R+D70R achieved a 13,680-fold preference of sp

59 Ecotin M84R+D70R bound to uPA at 50 pM, a 56,000-fold in

60 The structure of ecotin M84R-FXa (M84R-FXa) also reveals the structure of

61 ence, and the structures of the rhFXI370-607-ecotin mutant complexes were determined.

62 The presence of ecotin not only assisted in the crystallization of the e

63 ant p.G214R was inhibited poorly by eglin C, ecotin, or a CTRC-specific variant of SGPI-2, and it rea

64 nst bovine trypsin resulted in enrichment of ecotin phage but did not yield a consensus sequence.

65 Two ecotin phage display libraries were constructed to explo

66 to demonstrate that inhibition of PKal with ecotin-PKal delays alveolar apoptosis, adipocyte repleni

67 We used a variant of ecotin (ecotin-PKal), a macromolecular inhibitor of serine prote

68 Using biotinylated ecotin-PKal, we localized active PKal to connective tiss

69 The fact that the 60 s loop of ecotin plays different roles in binding to trypsin and u

70 tin-ecotin dimer interface, a larger primary ecotin-protease interface, and a smaller secondary ecoti

71 -protease interface, and a smaller secondary ecotin-protease interface.

72 he cross-linking reaction occurs between two ecotin proteins in a dimer complex.

73 A point mutation (M84R) in ecotin results in a 1.5 nM affinity for thrombin, 10(4)

74 through the use of wild-type and engineered ecotins results in inhibition of rat prostate differenti

75 sected to understand the structural basis of ecotin's broad inhibitory specificity and the role of th

76 egion-specific mutagenesis were preformed at ecotin's primary site P1 residue (84), the C-terminal di

77 rypsin and human uPA established the role of ecotin's secondary binding site in recognizing these hom

78 er secondary interaction between the partner ecotin subunit and the protease.

79 The ecotin surface loop that was redesigned is composed of r

80 acid substitution in the carboxy terminus of ecotin that places an engineered tyrosine within 5 A of

81 ue 99 and the C-terminus of thrombin contact ecotin through mixed polar and nonpolar interactions.

82 been developing the macromolecular inhibitor ecotin to be a "fold-specific" inhibitor that is selecti

83 using the three-dimensional structure of the ecotin-trypsin complex to guide combinatiorial design ef

84 cture of the previously determined wild-type ecotin-trypsin complex.

85 , we have solved the structure of two mutant ecotin-trypsin complexes and compared them to the struct

86 which has four alanine substitutions in the ecotin-trypsin secondary binding site, along with the M8

87 ses of the protein-protein interfaces in the ecotin-trypsin tetramer.

88 omplex shows minor structural changes in the ecotin-trypsin tetramer.

89 Ecotin undergoes a pronounced internal structural rearra

90 A monomeric ecotin variant (mEcotin) and a single-chain ecotin dimer

91 The structure of the tetrameric complex of ecotin variant M84R (M84R) with FXa has been determined

92 The contrast in the Ki values of the various ecotin variants towards bovine trypsin, rat trypsin and

93 ociation constants for monomeric and dimeric ecotin variants were determined with both trypsin and ch

94 rization constant (K(dim)) of wild-type (WT) ecotin was found to be picomolar by subunit exchange exp

95 Subsequently, the sequence from P5 to P2' in ecotin was mutated to the FXIa substrate sequence, and t

96 t position 84 in the primary binding site of ecotin, was generated for panning against uPA and rat tr

97 60X4 library, in which residues 67 to 70 of ecotin were randomized, was panned against rat and bovin

98 Allylglycine-containing ecotins were synthesized by in vitro translation of the

99 trypsin at 80 pM, a 12-fold improvement over ecotin wild-type (WT).

100 nteractions of the secondary binding site of ecotin with bovine trypsin, rat trypsin and human urokin

101 ine protease factor Xa (FXa) is inhibited by ecotin with picomolar affinity.

102 56,000-fold increase in binding compared to ecotin WT.

103 nM for trypsin, while the optimized mutant, ecotin Y69F, D70P, which was found using phage display t

104 The structure of the ecotin Y69F, D70P-trypsin complex shows minor structural