The Comparison of Ca2+/CaM-Independent
and Ca2+/CaM-Dependent Phosphorylation of Myosin
Light Chains by MLCK
Z. TANG, H. CHEN, J. YANG, S. DAI, Y. LIN
Department of Pharmacology, Dalian Medical University, Dalian,
China
Received October 20, 2004
Accepted December 14, 2004
On-line available February 16, 2005
Summary
The main regulatory mechanism of smooth muscle contraction
involves Ca2+/calmodulin (CaM)-dependent
phosphorylation of myosin (CDPM), by myosin light chain kinase
(MLCK). It is also known that the increase in intracellular Ca2+
and phosphorylation of myosin occurs within a short time under
physiological conditions, but the muscle tension may persist for
a longer period of time. However, the mechanism of this
phenomenon is still not clear. We hypothesize that MLCK also
phosphorylates myosin in a Ca2+/CaM-independent manner (CIPM).
The difference between CIPM and CDPM are as follows. Firstly,
the extent of CIPM by MLCK was temperature-independent, whereas
CDPM by MLCK was apparently decreasing with increasing
temperature. Secondly, in contrast to the decreased extent of
CDPM, the prolongation of incubation time did not decrease the
extent of CIPM. Thirdly, a high concentration of K+ influences
CIPM less than CDPM. Furthermore, the MLCK inhibitor ML-9
significantly inhibited CDPM by MLCK but not CIPM by MLCK.
Lastly, arachidonic acid selectively increased CIPM by MLCK but
not CDPM by MLCK. Finally, the activity of Mg2+-ATPase
of myosin followed the sequence as this: CDPM > CIPM >
unphosphorylated myosin. Our results revealed some primary
features of CIPM by MLCK.
Key words
Ca2+/CaM-independent phosphorylation • Ca2+/CaM-dependent
phosphorylation • Myosin • MLCK
|