Participation of Two Carboxyl Groups in Phosphodiester Hydrolysis. 2. A Kinetic, Isotopic, and 31P NMR Study of the Hydrolysis of a Phosphodiester with Carboxyl Groups Fixed in an Attack Conformation

1995 ◽  
Vol 117 (49) ◽  
pp. 12070-12077 ◽  
Author(s):  
Thomas C. Bruice ◽  
Andrei Blasko ◽  
Ramesh D. Arasasingham ◽  
Jang-Seob Kim
Keyword(s):  
31P Nmr ◽  
Carboxyl Groups ◽  
Nmr Study ◽  
Phosphodiester Hydrolysis ◽  
Hydrolysis Of

On the hydrolysis of dimethyl-H-phosphonate. An 18O-labelling and 31P-NMR study

Polyhedron ◽  
1998 ◽  
Vol 17 (4) ◽  
pp. 433-442 ◽  
Author(s):  
Michael C. Mitchell ◽  
Roger J. Taylor ◽  
Terence P. Kee
Keyword(s):  
31P Nmr ◽  
Nmr Study ◽  
Hydrolysis Of

scholarly journals Pseudomonas putida MPE, a manganese-dependent endonuclease of the binuclear metallophosphoesterase superfamily, incises single-strand DNA in two orientations to yield a mixture of 3′-PO4 and 3′-OH termini

2020 ◽  
Author(s):  
Shreya Ghosh ◽  
Anam Ejaz ◽  
Lucas Repeta ◽  
Stewart Shuman
Keyword(s):  
Pseudomonas Putida ◽  
Bound Water ◽  
Acid Catalyst ◽  
Nuclease Activity ◽  
Single Strand ◽  
Dna Endonuclease ◽  
Dependent Endonuclease ◽  
Leaving Group ◽  
Phosphodiester Hydrolysis ◽  
Hydrolysis Of

Abstract Pseudomonas putida MPE exemplifies a novel clade of manganese-dependent single-strand DNA endonuclease within the binuclear metallophosphoesterase superfamily. MPE is encoded within a widely conserved DNA repair operon. Via structure-guided mutagenesis, we identify His113 and His81 as essential for DNA nuclease activity, albeit inessential for hydrolysis of bis-p-nitrophenylphosphate. We propose that His113 contacts the scissile phosphodiester and serves as a general acid catalyst to expel the OH leaving group of the product strand. We find that MPE cleaves the 3′ and 5′ single-strands of tailed duplex DNAs and that MPE can sense and incise duplexes at sites of short mismatch bulges and opposite a nick. We show that MPE is an ambidextrous phosphodiesterase capable of hydrolyzing the ssDNA backbone in either orientation to generate a mixture of 3′-OH and 3′-PO4 cleavage products. The directionality of phosphodiester hydrolysis is dictated by the orientation of the water nucleophile vis-à-vis the OH leaving group, which must be near apical for the reaction to proceed. We propose that the MPE active site and metal-bound water nucleophile are invariant and the enzyme can bind the ssDNA productively in opposite orientations.

scholarly journals Accumulation of N-acyl-ethanolamine phospholipids in rat brains during post-decapitative ischemia: a 31P NMR study

1999 ◽  
Vol 40 (3) ◽  
pp. 515-521 ◽  
Author(s):  
Birthe Moesgaard ◽  
Jerzy W. Jaroszewski ◽  
Harald S. Hansen
Keyword(s):  
31P Nmr ◽  
Nmr Study
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