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

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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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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.}",

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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.

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VL - 6

SP - 1096

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JO - Nature Structural Biology

JF - Nature Structural Biology

IS - 12

ER -

A novel endonuclease mechanism directly visualized for I-PpoI

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