University of Arkansas

Other

Richard Drake

Cesar M Compadre

Barry Hurlburt

Tammy M Rechtin

Trenton Hinds

Alan D Elbein

Irena Patuszak

The present invention provides new site-specific HSV-thymidine kinase mutants with improved nucleoside analog metabolizing activity due to low or no thymidine phosphorylation ability. Also provided is a method of killing target cells using such mutants combined with a prodrug.

Mutated herpes simplex virus type 1 thymidine kinases and uses thereof

A new enzyme that phosphorylates GalNAc at position 1 to form GalNAc-&agr;-1P was purified &tilde;1275-fold from the cytosolic fraction of pig kidney, and the properties of the enzyme were determined. The kinase is specific for GalNAc as the phosphate acceptor and is inactive with GlcNAc, ManNAc, glucose, galactose, mannose, GalN, and GlcN. The native enzyme has a molecular mass of 48-51 kDa, and this enzyme is clearly separated from galactokinase by chromatography. The GalNAc kinase has a pH optimum between 8.5 and 9.0, and requires a divalent cation in the order Mg2&plus;>Mn2&plus;>Co2&plus;, with optimum Mg2&plus; concentration at &tilde;5 mM. The enzyme was most active with ATP as the phosphate donor, but slight activity was observed with ITP, acetyl-P, and phosphoenolpyruvate. Enzyme activity was highest in porcine and human kidney and porcine liver, and was low in most other tissues.

Liver GalNAc-1-phosphate kinase

Growing community of inventors

Little Rock, AR, United States of America

Richard Drake