Diabetes, Vol 47, Issue 10 1544-1548, Copyright © 1998 by American Diabetes Association
Distinguishing the type I and type II isozymes of hexokinase: the need for a reexamination of past practice
JE Wilson
Department of Biochemistry, Michigan State University, East Lansing 48824-1319, USA. wilsonj@pilot.msu.edu
The type I and type II isozymes of hexokinase coexist in insulin-sensitive
tissues, such as cardiac and skeletal muscle and adipose tissue. Based on
an early report that the purified type I isozyme was stable at 45 degrees C
whereas the purified type II isozyme was not, investigators in a number of
studies have used heat lability as a criterion for distinguishing these
isozymes in crude tissue extracts or subcellular fractions; that is,
activity lost after incubation at 45 degrees C was believed to be type II
while remaining activity was considered type I. This extrapolation is
dangerous because thermal lability can be markedly affected by the solvent
environment, including the presence or absence of other proteins. In the
present study, the rate of thermal inactivation of the type I isozyme has
been shown to vary by at least an order of magnitude in soluble and
particulate fractions prepared from rat heart and brain. Thus, the use of
thermal stability as a general criterion for identifying the type I isozyme
is invalid, and conclusions based on thermal inactivation as a means for
distinguishing the type I and type II isozymes need to be reconsidered.