Retention of Configuration in the Action of Human Plasma 3‘-Exonuclease on Oligo(deoxynucleoside phosphorothioate). A New Method for Assignment of Absolute Configuration at Phosphorus in Isotopomeric Deoxyadenosine 5‘-O-[18O]Phosphorothioate

A new method of analysis has allowed the exonucleolytic cleavage by human 3‘-exonuclease to be determined. Hydrolysis by human plasma 3‘-exonuclease proceeds with retention of configuration at phosphorus. The new method determines the sense of chirality at phosphorus in isotopomeric adenosine 5‘-O-[...

Full description

Saved in:
Bibliographic Details
Published in:Journal of the American Chemical Society 2002-05, Vol.124 (17), p.4623-4627
Main Authors: Koziołkiewicz, Maria, Owczarek, Alina, Wójcik, Marzena, Domański, Krzysztof, Guga, Piotr, Stec, Wojciech J
Format: Article
Language:English
Subjects:
Biological and medical sciences
Conformational dynamics in molecular biology
Deoxyadenosines - analogs & derivatives
Deoxyadenosines - chemistry
Deoxyadenosines - metabolism
Exonucleases - blood
Exonucleases - chemistry
Fundamental and applied biological sciences. Psychology
Humans
Molecular biophysics
Molecular Conformation
Phosphorus - chemistry
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Thionucleotides
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:A new method of analysis has allowed the exonucleolytic cleavage by human 3‘-exonuclease to be determined. Hydrolysis by human plasma 3‘-exonuclease proceeds with retention of configuration at phosphorus. The new method determines the sense of chirality at phosphorus in isotopomeric adenosine 5‘-O-[18O]phosphorothioates. This is based on stereospecific two-step conversion of the mono-thionucleotide into the corresponding deoxyadenosine 5‘-O-α-[18O]thiotriphosphate, followed by the use of terminal deoxyribonucleotidyl transferase and MALDI TOF mass spectrometry of the resulting elongated primer. Retention of configuration in the reaction of plasma 3‘-exonuclease implies a two-step mechanism with two displacements on phosphorus. Inversion at each step leads to overall retention.
ISSN:0002-7863
1520-5126
DOI:10.1021/ja017187u