This document summarizes the role of calcium and calmodulin in cellular signaling. It discusses how calcium acts as a regulatory second messenger by binding to proteins like calmodulin. Calmodulin has four calcium-binding sites that allow it to adopt an open confirmation and bind target sequences. Calmodulin regulates enzymes like calcium/calmodulin dependent protein kinases, myosin light chain kinases, and calcineurin. These enzymes play important roles in processes like muscle contraction, transcription, and neuronal signaling by phosphorylating or dephosphorylating target proteins in response to calcium signals.

1. M.Prasad Naidu
MSc Medical Biochemistry,
Ph.D.Research Scholar

2.  THE JOURNAL OF BIOLOGICAL CHEMISTRY
 Vol. 276, No. 4, Issue of January 26, pp. 2311–2312, 2001
 © 2001 by The American Society for Biochemistry and
Molecular Biology, Inc.
 Printed in U.S.A.

3.  Extracellular Ca2+ is necessary for muscle
contractions .
 Intracellular Ca2+ mediates a large number of cellular
responses with the high affinity and
specificity required for a regulatory second messenger.
 Ca2+ acts by binding to specific intracellular proteins,
which may be considered Ca2+ receptors .

4.  The class of proteins that binds Ca2+ with high affinity
and specificity now includes hundreds of members .
 Studies focused on calmodulin because it was
recognized as a regulator of many different target
enzymes .

5.  Calmodulin interactions with proteins are categorized
into six distinct classes based on Ca2+ -independent and
-dependent modes of binding and regulation .
 Calmodulin has four Ca2+ -binding sites with two in a
globular N-terminal domain separated by a flexible alfa-
helix from a C-terminal globular domain containing the
other two Ca2+ binding sites .

6.  In the presence of Ca2+ each domain adopts an open
conformation exposing a hydrophobic pocket that
renders calmodulin functional for binding to target
sequences .

7.  The process of complex formation includes sequential
interactions between the C-terminal hydrophobic
pocket with a hydrophobic residue in the target N-
terminal sequence followed by interactions between
the N-terminal globular domain with the C-terminal
sequence of the calmodulin-binding domain.

8.  Calmodulin thus collapses and wraps around the
peptide, resulting in the formation of a high affinity
complex.
 The high affinity binding of calmodulin to a target
sequence is only partly responsible for enzyme
activation as surface residues on calmodulin may
subsequently interact with other areas of the enzyme .

9.  Ca2+/calmodulin regulates
1. calmodulin-dependent protein kinases I, II, IV,
2. myosin light chain kinases,
3. calmodulin-dependent protein phosphatase,
4. calcineurin.

10.  Protein phosphatases play dynamic roles in diverse
cellular processess .
 Protein phosphatase IIB, or calcineurin, is a
serine/threonine protein phosphatase activated by
Ca2+/calmodulin and, thus,
couples Ca2+ signals to specific cellular responses via
protein dephosphorylation .

11.  Calcineurin was initially identified in neuronal
tissues, but it was quickly recognized that it had a
broad tissue distribution and a highly conserved
structure from yeast to man.
 The immunosuppressive drugs, cyclosporin A and
FK506 have provided tools to explore the roles of
calcineurin in diverse Ca2+dependent signaling
pathways .

12.  Calcineurin has role in regulating transcription during
development via dephosphorylation of the NF-AT
transcription complex.
 Ca2+ also regulates transcription through
phosphorylation of several transcription
factors, including CREB and MEF2, by a
Ca2+/calmodulindependent protein kinase cascade
involving three distinct kinases.

13.  Ca2+/calmodulin-dependent protein kinases I and IV
have broad but overlapping substrate specificities and
similar mechanisms of activation.
 Both kinases are phosphorylated by a third
kinase, which enhances Ca2+/calmodulin-dependent
activity .

14.  Another ubiquitous effector of Ca2+ signaling is the
multifunctional Ca2+/calmodulin-dependent protein
kinase II, which has
broad substrate specificity with substrates found in
nuclear, cytoskeletal, and membrane compartments of
cells .

15.  Multifunctional Ca2+/calmodulin-dependent protein
kinase II, which has biochemical property of
autophosphorylation mechanism that results in the
trapping of calmodulin on the kinase and conversion of
the enzyme to a Ca2+ independent form .

16.  The dynamic and spatial aspects of
autophosphorylation are an important element of
regulation, allowing it to respond to transient cellular
Ca2+ oscillations .
 Ca2+/calmodulin-dependent protein kinase II in the
neuronal synapse where its translocation and
activation regulate a number of proteins in the
postsynaptic cell.

17.  In contrast to the Ca2+/calmodulin-dependent protein
kinases and calcineurin that have broad substrate
specificities, myosin light chain kinases are dedicated
protein kinases for which the only known
physiological substrate is the regulatory light chain of
myosin II .

18.  Myosin light chain kinases have role cell spreading
and migration, cytokinesis, cell
adhesion, secretion, and cytoskeletal arrangements
that affect plasma membrane ion movements .

19.  Smooth muscle sarcoplasm contain a myosin light
chain kinase that is calcium dependent .
 The Ca 2+ activation of myosin light chain kinase
requires binding of calmodulin .4Ca2+ to its Kinase
subunit .
 Calmodulin .4Ca2+ acivated light chain kinase
phosphorylates the p – light chain , which then ceases
to inhibit the myosin - F – actin interaction , the
contraction cycle then begins.

20. Enzymes regulated by calcium or
calmodulin
 Adenylyl cyclase , guanylyl cyclase ,
 Cyclic nucleotide phosphodiesterase ,
 Calcium dependent protein kinase , phospho protein
phosphatse 2 B,
 Glycerol 3 phosphate dehydrogenase , Phosphorylase kinase ,
glycogen synthase ,
 Pyruvate carboxylase , pyruvate kinase ,
 Ca2+ Mg2+ ATPase ,
 Phospholipase A2 , Ca2+ phospholipid dependent protein
kinase