ライフサイエンスコーパス: extein

1 rsors during maturation of the host protein (extein).

2 itant ligation of the flanking polypeptides (exteins).

3 ocess ligate the flanking protein sequences (exteins).

4 e joining of the flanking protein sequences (exteins).

5 rmophilic Pyrococcus abyssi PolII intein and extein.

6 s cleavage of the intein from its N-terminal extein.

7 ptide bond cleavage between the intein and C-extein.

8 intein, is removed from a host protein, the extein.

9 in level from the flanking host protein, the exteins.

10 oncomitant with the specific ligation of the exteins.

11 rm a native peptide bond between the ligated exteins.

12 y 100 amino acids long, fused to appropriate exteins.

13 er native conditions was achieved using mini exteins.

14 five mutations at the first residue of the C-extein (+1), and describe molecular properties that may

15 rmation of a thioester linkage between the N-extein and intein.

16 tions between distant residues in the native exteins and the intein, in three-dimensional space.

17 emselves out of precursor proteins while the exteins are ligated together.

18 o the formation of new proteins in which the exteins are linked directly by a peptide bond.

19 o the formation of new proteins in which the exteins are linked directly by a peptide bond.

20 ed and the remaining two flanking sequences (exteins) are joined.

21 eir ligation junction (called local N- and C-exteins) are strongly preferred, while other sequences c

22 ols or Cys despite an ester linkage at the C-extein branch point, and (d) an absolute requirement for

23 ctrostatic interactions between the N- and C-exteins but is reduced by elevated temperature.

24 t ligation of the flanking polypeptides, the exteins, by a peptide bond.

25 g of the flanking polypeptide sequences, the exteins, by a peptide bond.

26 of the flanking sequences, the extein-N and extein-C parts, thereby reconstituting the host protein.

27 -splicing junction in both N- and C-terminal exteins can accelerate the N-terminal cleavage rate by >

28 netic analysis of the relationship between C-extein composition and split intein activity.

29 e their excision from flanking polypeptides (exteins) concomitant with extein ligation to produce a m

30 the intein, from flanking polypeptides, the exteins, concomitant with the specific ligation of the e

31 stoc punctiforme Npu DnaE intein after the C-extein cysteine nucleophile (Cys+1) was mutated to serin

32 used naturally split inteins suffer from an "extein dependence," whereby residues surrounding the spl

33 es ultrafast DnaE split inteins with minimal extein dependence.

34 The existence and the magnitude of extein effects require consideration for maximizing the

35 rsor proteins while ligating the surrounding extein fragments with a native peptide bond.

36 that inteins may act as switches to control extein function.

37 in splicing precursor, corresponding to an N-extein fusion of the Mxe GyrA intein.

38 identity of the C-terminal residue of the N-extein has less influence on the cleavage reaction than

39 inteins present in the same location within extein homologs from different organisms are very closel

40 for the (-1) scissile peptide bond at the N-extein-intein junction was found to be approximately 12

41 n the intein and the split-host protein, the exteins, is largely unknown.

42 ins, from flanking polypeptide sequences, or exteins, leading to the formation of new proteins in whi

43 Applying this assay to randomized extein libraries, we show that the nature of the extein

44 s and reactive nitrogen species inhibit SufB extein ligation by forcing either precursor accumulation

45 king polypeptides (exteins) concomitant with extein ligation to produce a mature host protein.

46 ion of defined regions of a protein prior to extein ligation, generating partially labeled proteins f

47 The exteins might thereby serve as an environmental sensor,

48 tant ligation of the flanking sequences, the extein-N and extein-C parts, thereby reconstituting the

49 In this non-canonical pathway, the C-extein nucleophile attacks a peptide bond at the N-termi

50 the intein for full activity, as can remote extein point mutations.

51 ein is compatible with any amino acid in the extein position adjacent to the N-terminal splice juncti

52 -extein specificity and only two important C-extein positions.

53 hen FKBP and FRB occupied one or both of the extein positions.

54 ld binds to two inactive split intein/enzyme extein protein fragments leading to intein fragment comp

55 that were as fast or faster than the native extein, refuting past assumptions that the naturally sel

56 udy, we investigated the roles of the last N-extein residue (-1 residue) and the intein penultimate r

57 lar insight into previous claims that the +1 extein residue affects intein catalysis.

58 a proton transfer from the first C-terminal extein residue to a conserved aspartate, which synchroni

59 tic residues are constrained by the second C-extein residue, likely forcing them into an active confo

60 riplet (WCT) and a Thr or Ser as the first C-extein residue.

61 niprecursor (VMA29) containing 10 N-terminal extein residues and 4 C-terminal extein residues.

62 the Ssp DnaE intein containing five native N-extein residues and maltose binding protein as the N-ext

63 conditions and also in the absence of native extein residues flanking the intein.

64 N-terminal extein residues and 4 C-terminal extein residues.

65 ue with alanine and mutation of the native C-extein residues.

66 ing a hexahistidine sequence as the N- and C-exteins, respectively.

67 y sequence similarity to only the selected C-extein sequence.

68 y may be affected by both the intein and the extein sequence.

69 mptions that the naturally selected flanking extein sequences are optimal for splicing.

70 with concomitant linkage of the two flanking extein sequences by a native peptide bond.

71 The extein sequences immediately flanking the intein affect

72 ution of the split intein, were fused to the extein sequences of the split intein halves.

73 ith the concomitant ligation of the flanking extein sequences to yield a new polypeptide.

74 The novel selected extein sequences were sufficient to promote splicing in

75 In this study, genetic selection identified extein sequences with Ser+1 that enabled the Npu DnaE in

76 together with the ligation of the flanking "extein" sequences.

77 ences than previously thought, with little N-extein specificity and only two important C-extein posit

78 in libraries, we show that the nature of the extein substrate bordering the intein can profoundly inf

79 ysis showed splicing rates with the selected exteins that were as fast or faster than the native exte

80 sion from flanking polypeptide sequences, or exteins, thereby leading to the formation of new protein

81 eins and ligate their flanking polypeptides (exteins) through a multistep chemical reaction.

82 t 100 amino acids each, fused to appropriate exteins, was recently derived from the Mycobacterium tub

83 ltose-binding protein (MBP) and a His-tag as exteins were expressed from a constitutive promoter afte

84 the intein, from flanking polypeptides, the exteins, which are concomitantly joined by a peptide bon

85 This partnership between the intein and its exteins, which implies coevolution of the parasitic inte

86 rikoshii is strongly regulated by the native exteins, which lock the intein in an inactive state.

87 esidues and maltose binding protein as the N-extein with the C-terminal Ssp DnaE intein splicing doma

88 concatenation of the flanking polypeptides (exteins) with a native peptide bond.