CST Guide:

Section I: Research Areas
Calcium pump
protein ATP2A2/
SERCA2 is widely
expressed in
many cell lines.
Calcium Channels and Pumps
Maintaining proper calcium concentrations within the cell is critical for effective cell signaling and
requires a variety of channels and pumps to transport calcium ions across intracellular and plasma
membranes. Ion channels move calcium ions into the cell or out from intracellular storage compartments
(with the gradient), effectively raising cytoplasmic calcium concentrations. The three types of
calcium ion channels are broadly classified by their ability to open in response to a ligand, second
messenger, or membrane potential (voltage-dependent calcium channels; VDCC). For example, the IP3
receptor requires the second messenger inositol 1,4,5-triphosphate (IP3) for activation and is located
on the endoplasmic reticulum (ER) where it regulates release of intracellular calcium stores.
As cytoplasmic calcium concentrations rise, calcium can be transported back outside the cell or into
storage within the sarcoplasmic reticulum or ER by calcium pumps. Calcium pump proteins are calcium-
ATPases that use the energy of ATP hydrolysis to retrotransport calcium across plasma or ER membranes,
thus maintaining the calcium gradient necessary for rapid signaling. For example, the calcium
pump ATP2A2/SERCA2 is responsible for regulating calcium transport across sarcoplasmic reticulum and
ER membranes, and its activity can be regulated through a variety of post-translational modifications.
ATP2A2/SERCA2 Antibody #4388: WB analysis
of extracts from various cell lines using #4388.
Lanes
1. Hep G2
2. RD
3. C2C12
4. Jurkat
5. NIH/3T3
6. PC-12
IP3 receptor, a calcium ion channel activated by
second messengers, is expressed in brain tissue.
IP3 Receptor 1 (D53A5) Rabbit mAb #8568: WB analysis of extracts from mouse and
rat brain using #8568.
82 For Research Use Only. Not For Use in Diagnostic Procedures. See pages 302 & 303 for Pathway Diagrams, Application, and Reactivity keys.
kDa
200
140
100
80
60
50
40
kDa
200
140
100
1 2
IP3
Receptor 1
Lanes
1. mouse brain
2. rat brain
Mitochondrial Calcium Uniporter
The mitochondrial calcium uniporter (MCU) is a calcium channel specifically located within the
mitochondrial inner membrane. Mitochondrial calcium uniporter regulator 1 (MCUR1) is a multi-pass,
transmembrane protein that directly interacts with MCU and plays an essential role in the regulation of
calcium uptake and maintenance of mitochondrial calcium homeostasis. Regulation of MCU by MCUR1
may be critical for a variety of cellular functions, including signal transduction, bioenergetics, and cell
death and survival.
Mitochondrial calcium uniporter
is expressed in many cell lines.
MCUR1 Antibody #13706: WB analysis of extracts from
various tissues and cell lines using #13706 (upper) and
β-Actin (D6A8) Rabbit mAb #8457 (lower).
1 2 3 4 5 6
kDa
60
50
40
30
20
50
40
ATP2A2/
SERCA2
1 2 3 4 5 6
MCUR1
β-Actin
Lanes
1. mouse kidney
2. mouse testis
3. human kidney
4. rat kidney
5. C6
6. Neuro-2a
Select Reviews
Bublitz, M., Musgaard, M., Poulsen, H., et al. (2013) J. Biol. Chem. 288, 10759–10765. • Freeley, M., Kelleher, D., Long, A.
(2011) Cell. Signal. 23, 753–762. • Newton, A.C. (2010) Am. J. Physiol. Endocrinol. Metab. 298, 395–402. • Patron, M.,
Raffaello, A., Granatiero, V. et al. (2013) J. Biol. Chem. 288, 10750–10758. • Rossi, A.M., Tovey, S.C., Rahman, T., et al.
(2012) Biochim. Biophys. Acta. 1820, 1214–1227. • Yáñez, M., Gil-Longo, J., and Campos-Toimil, M. (2012) Adv. Exp. Med.
Biol. 740, 461–482. • Yang, Y.R., Follo, M.Y., Cocco, L., and Suh, P.G. (2013) Adv. Biol. Regul. 53, 232–241.
Commonly Studied Calcium, cAMP, and Lipid Signaling Targets
Target M P S Target M P S Target M P S
β1-Adrenergic Receptor • PDE5
• Phospho-Phospholamban
(Ser16/Thr17)
•
AKAP1 •
PIP4K2A •
AKAP5 •
PIP4K2B • PLCβ3 • •
Annexin A1 • • PIP5K1A • Phospho-PLCβ3 •
(Ser537)
Annexin A2 •
PIP5K1C •
Phospho-PLCβ3
Annexin A7 • PKA C-α • • •
•
(Ser1105)
ApoA1 •
Phospho-PKA C-α • •
PLCγ1 • •
ApoA4 •
(Thr197)
Phospho-PLCγ1
ApoA5 •
PKA RI-α/β •
• •
(Tyr783)
ApoM •
Phospho-PKC (pan) •
Phospho-PLCγ1 • •
(βII Ser660)
ASM
•
(Ser1248)
Phospho-PKC (pan)
ATP2A1/SERCA1 • •
•
PLCγ2
•
(γ Thr514)
ATP2A2/SERCA2 • •
Phospho-PLCγ2
Phospho-PKC (pan) •
•
Pan-Calcineurin A •
(Tyr759)
(ζ Thr410)
Calmodulin •
Phospho-PLCγ2
PKCα
•
•
(Tyr1217)
Calumenin •
Phospho-PKCα/β II •
PLD1
CBARA1/MICU1 •
•
(Thr638/641)
Phospho-PLD1 (Thr147)
CFTR
• PKCδ • •
•
Phospho-PLD1 (Ser561)
Choline Kinase α •
Phospho-PKCδ (Tyr311) •
•
PLD2
cPLA2 • • Phospho-PKCδ (Thr505) •
•
PRK2
Phospho-cPLA2 • Phospho-PKCδ/θ •
• •
(Ser505)
(Ser643/676)
PKA RI-α •
Cyclic AMP •
PKCε • • RyR1 •
DAG Lipase β •
PKCθ • • S100A1 •
Gα (pan) • Phospho-PKCθ (Thr538) • S100A4 • •
Gα (z)
• PKCζ • • • S100A6 •
Gα (i)
• Phospho-PKCζ/λ • S100A10 •
Gα (o)
•
(Thr410/403)
S100B •
Gelsolin • • •
PKD/PKCµ • S100P
• •
INPP4b • •
Phospho-PKD/PKCµ • nSMase1 •
(Ser744/748)
IP3 Receptor • •
SPHK1 • •
Phospho-PKD/PKCµ
Phospho-IP3 Receptor • •
• STIM1 • •
(Ser916)
(Ser1756)
STIM2
•
PKD2
MARCKS •
• •
TGM2 •
PKD3/PKCν
Phospho-MARCKS • •
•
TRPV3
•
(Ser152/156)
Phospho-PRK1 •
TSPO
•
(Thr774)/PRK2 (Thr816)
Phospho-MARCKS •
WFS1
(Ser167/Ser170)
Phospholamban •
• •
NIPSNAP1 •
Select Citations:
Volk, L. et al. (2013) PKM-zeta is not required for hippocampal
synaptic plasticity, learning and memory. Nature
493, 420–423.
Paul, S. et al. (2014) T cell receptor signals to NF-kappaB
are transmitted by a cytosolic p62-Bcl10-Malt1-IKK signalosome.
Sci. Signal. 7, ra45.
Dusaban, S.S. et al. (2013) Phospholipase C epsilon links
G protein-coupled receptor activation to inflammatory
astrocytic responses. Proc. Natl. Acad. Sci. USA 110,
3609–3614.
Xiang, S.Y. et al. (2013) PLCepsilon, PKD1, and SSH1L
transduce RhoA signaling to protect mitochondria from
oxidative stress in the heart. Sci. Signal. 6, ra108.
Varsano, T. et al. (2013) Inhibition of melanoma growth by
small molecules that promote the mitochondrial localization
of ATF2. Clin. Cancer Res. 19, 2710–2722.
Ke, G. et al. (2013) MiR-181a confers resistance of cervical
cancer to radiation therapy through targeting the proapoptotic
PRKCD gene. Oncogene 32, 3019–3027.
Stumpf, C.R. et al. (2013) The translational landscape of the
mammalian cell cycle. Mol. Cell. 52, 574–582.
Gobbi, G. et al. (2013) Proplatelet generation in the mouse
requires PKCepsilon-dependent RhoA inhibition. Blood 122,
1305–1311.
Tuszynski, M.H. et al. (2012) Concepts and methods for
the study of axonal regeneration in the CNS. Neuron 7,
777–791.
Qu, Y. et al. (2012) Phosphorylation of NLRC4 is critical for
inflammasome activation. Nature 490, 539–542.
chapter 02: Signaling
These protein targets represent key
nodes within calcium, cAMP, and lipid
signaling pathways and are commonly
studied in calcium, cAMP, and lipid
signaling research. Primary antibodies,
antibody conjugates, and antibody
sampler kits containing these targets
are available from CST.
Listing as of September 2014. See our
website for current product information.
M Monoclonal Antibody
P Polyclonal Antibody
S SignalSilence ® siRNA
58
2012–2014 citations
CST antibodies for PKC have been
cited over 58 times in high-impact, peerreviewed
publications from the global
research community.
www.cellsignal.com/cstcalcium 83