A novel endonuclease mechanism directly visualized for I-PpoI

E. A. Galburt; B. Chevalier; W. Tang; M. S. Jurica; K. E. Flick; Jr Monnat; B. L. Stoddard

doi:10.1038/70027

A novel endonuclease mechanism directly visualized for I-PpoI

E. A. Galburt
, B. Chevalier
, W. Tang
, M. S. Jurica
, K. E. Flick
, Jr Monnat
, B. L. Stoddard

Research output: Contribution to journal › Article › peer-review

104 Scopus citations

Abstract

A novel mechanism of DNA endonucleolytic cleavage has been visualized for the homing endonuclease I-PpoI by trapping the uncleaved enzyme-substrate complex and comparing it to the previously visualized product complex. This enzyme employs a unique single metal mechanism. A magnesium ion is coordinated by an asparagine residue and two DNA oxygen atoms and stabilizes the phosphoanion transition state and the 3'oxygen leaving group. A hydrolytic water molecule is activated by a histidine residue for an in-line attack on the scissile phosphate. A strained enzyme-substrate-metal complex is formed before cleavage, then relaxed during the reaction.

Original language	English
Pages (from-to)	1096-1099
Number of pages	4
Journal	Nature Structural Biology
Volume	6
Issue number	12
DOIs	https://doi.org/10.1038/70027
State	Published - 1999

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title = "A novel endonuclease mechanism directly visualized for I-PpoI",

abstract = "A novel mechanism of DNA endonucleolytic cleavage has been visualized for the homing endonuclease I-PpoI by trapping the uncleaved enzyme-substrate complex and comparing it to the previously visualized product complex. This enzyme employs a unique single metal mechanism. A magnesium ion is coordinated by an asparagine residue and two DNA oxygen atoms and stabilizes the phosphoanion transition state and the 3'oxygen leaving group. A hydrolytic water molecule is activated by a histidine residue for an in-line attack on the scissile phosphate. A strained enzyme-substrate-metal complex is formed before cleavage, then relaxed during the reaction.",

author = "Galburt, \{E. A.\} and B. Chevalier and W. Tang and Jurica, \{M. S.\} and Flick, \{K. E.\} and Jr Monnat and Stoddard, \{B. L.\}",

year = "1999",

doi = "10.1038/70027",

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volume = "6",

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journal = "Nature Structural Biology",

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T1 - A novel endonuclease mechanism directly visualized for I-PpoI

AU - Galburt, E. A.

AU - Chevalier, B.

AU - Tang, W.

AU - Jurica, M. S.

AU - Flick, K. E.

AU - Monnat, Jr

AU - Stoddard, B. L.

PY - 1999

Y1 - 1999

N2 - A novel mechanism of DNA endonucleolytic cleavage has been visualized for the homing endonuclease I-PpoI by trapping the uncleaved enzyme-substrate complex and comparing it to the previously visualized product complex. This enzyme employs a unique single metal mechanism. A magnesium ion is coordinated by an asparagine residue and two DNA oxygen atoms and stabilizes the phosphoanion transition state and the 3'oxygen leaving group. A hydrolytic water molecule is activated by a histidine residue for an in-line attack on the scissile phosphate. A strained enzyme-substrate-metal complex is formed before cleavage, then relaxed during the reaction.

AB - A novel mechanism of DNA endonucleolytic cleavage has been visualized for the homing endonuclease I-PpoI by trapping the uncleaved enzyme-substrate complex and comparing it to the previously visualized product complex. This enzyme employs a unique single metal mechanism. A magnesium ion is coordinated by an asparagine residue and two DNA oxygen atoms and stabilizes the phosphoanion transition state and the 3'oxygen leaving group. A hydrolytic water molecule is activated by a histidine residue for an in-line attack on the scissile phosphate. A strained enzyme-substrate-metal complex is formed before cleavage, then relaxed during the reaction.

UR - https://www.scopus.com/pages/publications/0032708112

U2 - 10.1038/70027

DO - 10.1038/70027

M3 - Article

C2 - 10581547

AN - SCOPUS:0032708112

SN - 1072-8368

VL - 6

SP - 1096

EP - 1099

JO - Nature Structural Biology

JF - Nature Structural Biology

IS - 12

ER -

A novel endonuclease mechanism directly visualized for I-PpoI

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