| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
and Src tyrosine kinase
1 Department of Physiology and Biophysics, 2 Divisions of Cardiology and Nephrology, Department of Medicine, 3 Department of Biochemistry, 4 Core Proteomics Laboratory, and 5 Department of Pharmacology and Toxicology, University of Louisville, and 6 Veterans Affairs Medical Center, Louisville, Kentucky 40202
 |
ABSTRACT |
|---|
 TOP
 ABSTRACT
 INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
Our laboratory has conducted
multiple functional proteomic analyses to characterize the components
of protein kinase C (PKC)
cardioprotective signaling complexes and
found that activation of PKC induces dynamic modulation of these
complexes. In addition, it is known that signal transduction within a
complex involves the formation of modules, one of which has been shown
to include PKC and Src tyrosine kinase in the rabbit heart. However,
the cellular mechanisms that define the assembly of PKC modules
remain largely unknown. To address this issue, the interactions between PKC and Src were studied. We used recombinant proteins of wild-type PKC (PKC-WT) and open conformation mutants of the kinase
(PKC-AE5 and PKC-AN59), the regulatory and catalytic domains of
PKC, along with glutathione-S-transferase (GST) fusion proteins of Src (GST-Src) and two domains of Src (GST-SH2 and GST-SH3). GST pulldown assays demonstrated that Src and PKC are binding partners and that the interaction between PKC and Src appears to involve multiple sites. This finding was supported for endogenous PKC and
Src in the murine heart using immunofluorescence-based confocal microscopy and coimmunoprecipitation. Furthermore, PKC-WT and GST-Src interactions were significantly enhanced in the presence of
phosphatidyl-L-serine, an activator of PKC, indicating that Src favors interaction with activated PKC. This finding was
confirmed when the PKC-WT was replaced with PKC-AE5 or
PKC-AN59, demonstrating that the conformation of PKC is a
critical determinant of its interactions with Src. Together, these
results illustrate that formation of a signaling module between PKC
and Src involves specific domains within the two molecules and is
governed by the molecular conformation of PKC.
protein-protein interactions; cardioprotection; signaling complex; functional proteomics
| |
INTRODUCTION |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
MOUNTING EVIDENCE SUPPORTS the
notion that the assembly of multiprotein signaling complexes is a means
by which the cell accomplishes signal transduction (4, 8,
20). The exact manner in which a complex of proteins might be
regulated to confer distinct cellular responses (and ultimately organ
phenotypes, such as cardioprotection) remains unknown. One theory
provides that signaling modules may be assembled within the complexes
and that these modules may in turn direct subcellular tasks (24,
25). In an effort to understand, on a molecular level, the
mechanisms that influence the formation of modules, the present study
characterized the module containing protein kinase C (PKC) and Src.
The multifarious role of PKC in cellular signaling is well
established (Refs. 1, 3, 5,
10, 12, 14-16,
19-21, 22a, 23, and
26). It has been shown to be activated by a host of
stimuli and is known to participate in protection against
ischemic damage in numerous organs, especially the heart
(5, 19, 20). During cardioprotection, different members of
the PKC complex exhibit altered levels of association with the
complex and have been shown to undergo posttranslational modification
within this complex (19). Despite this, the principal
molecular mechanisms contributing to the assembly of the PKC complex
have never been investigated. In the cytosol, it is known that PKC
resides in a closed conformation, which is maintained by occupation of
its active site by a pseudosubstrate domain (14, 15). Upon
activation and translocation, the pseudosubstrate domain is released,
and PKC assumes an active and open conformation (14,
15). Accordingly, we tested the hypothesis that the molecular conformation of PKC plays a key role in cardioprotective PKC module assembly. Previous studies by our laboratory and others (Refs.
1, 21, 22a, and 25)
have indicated that PKC isoforms and Src family members may interact to
perform signaling tasks, and activation of PKC results in enhanced
activity of tyrosine kinases in the protection against ischemic
injury (Refs. 6, 7, 21,
22a, 23, and 25). Therefore, we
examined the PKC-Src module in the present study. With the use of
pharmacological stimulation, site-directed mutagenesis, in vitro
affinity assays, coimmunoprecipitation, and confocal microscopy, the
present investigation demonstrates that the molecular conformation of
PKC dramatically alters its interaction with Src tyrosine kinase and
serves as a major regulatory index of the association of PKC with Src.
| |
METHODS |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
Recombinant proteins and reagents.
Two mutants of PKC were used in the present study, both of which
render the kinase in an open conformation (PKC-AE5 and PKC-AN59).
The cDNAs of wild-type Src and the SH2 and SH3 domains of Src were
cloned into the pAcGHLT vector, expressed in the baculovirus system,
and purified to generate glutathione-S-transferase (GST) fusion proteins (BD PharMingen). Cold or
[35S]Met-labeled recombinant proteins of PKC-WT,
PKC-AE5, PKC-AN59, and the catalytic (PKC-CHA) and regulatory
(PKC-RHA) domains of PKC were also made via in vitro
transcription and translation using the TNT Quick-Coupled rabbit
reticulocyte lysate system (Promega). PKC and GST-Src were verified
to retain their kinase activity (data not shown). The lipid sources
were as follows: phosphatidyl-L-serine (PS; BioChemika) and
phorbol 12-myristate 13-acetate (PMA; Sigma).
Assessing protein-protein interactions via GST-based affinity pulldown assays. GST recombinant protein-based affinity pulldown assays were performed as previously reported (19). Briefly, GST recombinant proteins were immobilized on GST beads, mixed with either recombinant proteins of interest or cardiac tissue homogenates, and incubated in binding buffer [0.5% Triton X-100, 20 mM Tris · HCl (pH 7.4), 1 mM EDTA, 1 mM EGTA, 0.2 mM sodium orthovanadate, and 1× proteinase cocktail (Roche)]. The GST-protein pellets were washed, resolved by SDS-PAGE, and analyzed via immunoblotting with antibodies against the corresponding proteins or via autoradiography. To determine nonspecific binding, parallel reactions were conducted using equal molar amounts of GST-null proteins in place of the tested GST fusion proteins.
Coimmunoprecipitations.
PKC complexes were isolated from the hearts of
PKC-cardioprotected mice and nontransgenic controls as described
(19). Immunoblotting for Src was carried out using
antibodies from Santa Cruz. Coimmunoprecipitation for Src was done to
assess interactions with PKC-CHA and PKC-RHA as previously
described (17, 19).
Immunofluorescence-based confocal microscopy.
Sections of mouse left ventricular tissue were immunostained as
previously described (9, 11). Briefly, hearts were
excised, rinsed in PBS, fixed in 10% neutral-buffered formalin,
dehydrated in alcohol, embedded in paraffin, sectioned at a thickness
of 6 µm, and mounted on gelatin-coated slides. The sections were blocked in PBS containing 0.1% Triton X-100, 7% goat serum, and 5%
nonfat dry milk for 10 min at 37°C and then again with mouse IgG
Blocking Reagent (FMK-2201, Vector Laboratories) for an additional 1 h at RT. Antibody dilutions were as follows: PKC, 1:100 (BD Transduction Laboratories) overnight at 4°C; and Src, 1:100 (Santa Cruz and Biosource) for 2 h at 37°C. Sections were washed and incubated with secondary antibodies (TRITC-conjugated goat anti-rabbit and anti-mouse IgG antibodies at 1:200 dilution) for 1 h at
37°C. Negative controls were performed by omission of primary
antibodies. Optical sections were obtained with a Zeiss LSM510 inverted
confocal scanning laser microscope equipped with Argon/HeNe1 lasers
with excitation wavelengths appropriate for single channel scanning in
the individual tracks, FITC/TRITC, respectively. All experiments were
performed in triplicate, and all images were recorded within 24 h
of each other and analyzed with Adobe PhotoShop 5.5 software.
| |
RESULTS |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
Src favors interaction with active PKC.
The role of molecular conformation in the interaction of PKC with
Src tyrosine kinase was first investigated by using the lipid PS to
activate PKC-WT. As shown in Fig.
1A, the interaction of PKC
with Src was enhanced when PKC was activated with PS, whereas the
addition of PMA did not appear to increase the binding between these
two molecules relative to that observed in the absence of lipid
activators. Furthermore, the addition of PS and PMA did not appear to
induce any synergistic increase in association between Src and PKC
beyond that observed in the presence of PS alone (Fig.
1A).
|
. PKC-WT was found to interact with
GST-Src (Fig. 1B), a phenomenon that previous studies (25) have indicated is specific in that it varies directly
with the concentration of PKC protein. However, the binding of
PKC to Src was enhanced when PKC was rendered in its open
conformation by a mutation that prevents the binding of the
pseudosubstrate domain to the catalytic domain (16) (Fig.
1B), indicating that the interaction between PKC and Src
favors the open and active conformation of PKC.
Interaction between PKC and Src involves multiple domains on
Src.
To examine whether the binding of PKC to Src was regulated by
multiple domains on Src, we determined the interaction of GST-SH2 and
GST-SH3 fusion proteins with PKC. We observed that in vitro translated PKC bound to both the SH2 and SH3 domains of Src and that
it did so with a similar affinity (Fig. 1C). Importantly, this finding was confirmed using particulate fractions of mouse hearts
containing endogenous PKC (Fig. 1C).
Src preferentially interacts with the regulatory domain of PKC.
To characterize the domain of PKC that was responsible for its
interaction with Src, we used recombinant proteins of the regulatory
and catalytic regions of PKC. Recombinant Src was incubated with
[35S]Met-labeled PKC-CHA or PKC-RHA, and
coimmunoprecipitation for Src was performed. As shown in Fig.
1D, the interaction between the PKC-RHA and Src is much
stronger than that seen between the PKC-CHA and Src. Primary
sequence analysis of the two domains indicates that the ratio of
methionines in recombinant PKC-CHA relative to those in PKC-RHA
is 5:2. Therefore, a given amount of PKC-CHA protein will contain
2.5 times the number of methionines as the same amount of PKC-RHA
protein. Because the signal recorded for the CHA/RHA domains in Fig.
1D is based on autoradiography, one would anticipate that
the resultant signal from two equal amounts of PKC-CHA and
PKC-RHA protein would be ~2.5 times higher in the case of the
PKC-CHA. Such a positive control is seen by direct loading of
PKC-RHA and PKC-CHA (lanes 7 and 8,
respectively). This consideration suggests that the preference of Src
for PKC-RHA over PKC-CHA is actually higher than it appears,
because an equal amount of autoradiographic signal from PKC-CHA and
PKC-RHA would require less PKC-CHA than PKC-RHA protein.
Distribution of PKC and Src in the mouse heart.
Having shown in vitro that Src preferentially interacts
with the open conformation of PKC, we sought to confirm these
findings in vivo. We used a line of cardioprotected PKC mice
(16) to characterize the association of Src with PKC
complexes via coimmunoprecipitation and immunoblotting. As shown in
Fig. 2A, PKC-mediated
cardioprotection is clearly associated with increased localization of
Src to PKC complexes (increased total expression of Src was also
observed via Western blotting; data not shown). While Fig.
2A shows interaction of PKC and Src in the membrane
fraction, this interaction was also observed at numerous other
subcellular locations (data not shown). Next, we determined the effect
of PKC activation on PKC and Src distribution using
immunofluorescence microscopy. PKC-mediated cardioprotection was
accompanied by increased distribution of PKC to numerous subcellular
compartments (Fig. 2B). Furthermore, Src appeared to
localize to numerous areas of the cell and lacked specific
sequestration to any one particular subcellular location. These
findings were confirmed using multiple antibodies against different
epitopes of PKC and Src. A nearly identical pattern was seen for
both kinases with multiple antibodies, suggesting that the confocal
images obtained are truly representative of the subcellular
distribution of the respective molecule. Most importantly, there was a
strong correlation between the distribution of these two kinases
(PKC and Src), as indicated by their respective immunofluorescent
images, suggesting that the biochemical data we obtained via
coimmunoprecipitation are representative of increased association of
these two molecules at numerous subcellular locations. This finding was
further verified by independently examining localization of PKC and
Src to various subcellular locations (data not shown).
|
| |
DISCUSSION |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
In this study, we tested the hypothesis that the assembly of
signaling modules is dependent on the molecular conformation of the
proteins involved. We used recombinant proteins to demonstrate that
PKC and Src tyrosine kinase interact with a higher affinity when
PKC is in an active and open conformation. The particulate localization of these two molecules was confirmed in myocardial tissue
through coimmunoprecipitation, GST pulldown assays, and fluorescence-based confocal microscopy.
There are a number of important findings in this investigation. First,
it was shown that the formation of signaling modules is reliant in part
on the molecular conformation of the molecules within them. Second, it
was discovered that Src binds preferentially to the open conformation
of PKC, supporting the notion that the interaction of these two
molecules facilitates signal transduction (i.e., Src is more inclined
to associate with the activated PKC). Third, the interaction between
PKC and Src was found to favor the regulatory domain of PKC and
to involve multiple sites on Src. Finally, confocal microscopy and
coimmunoprecipitation were used to show that PKC and Src interact in
multiple particulate subcellular locations in the murine myocardium.
Role of PS and PMA.
While numerous studies have recently identified the colocalization of
multiple molecules as a means for signal transduction (2, 4, 8,
19), the manner in which the association of these molecules with
each other is regulated by molecular conformation is not understood.
The finding that addition of PS, but not PMA, enhanced the interaction
between PKC and Src is particularly interesting. The ability of
diacylglycerol (DAG) (phorbol esters/PMA) to elicit translocation of
PKC to the membrane has been extensively characterized (14,
15). This translocation is generally concomitant with activation
of the kinase, an occurrence regulated by PS (14, 15).
Importantly, the role of DAG/PMA appears to be exclusively for the
localization of PKC to the lipid bilayer membrane, and not directly
for the activation of PKC, in that the translocation seen in response
to PMA occurs in the absence of a conformational change in the
substrate-binding region of PKC (14, 15). In the present
study, the in vitro analysis obviates the question of translocation and
therefore places focus on the role of molecular configuration on the
interaction of PKC with Src. Because our previous studies
(25) have shown that PKC and Src interact in the
particulate region and not the cytosolic region, we sought in the
present study to understand if this was merely due to increased abundance of the two molecules in particulate regions or also dependent
on altered molecular conformation.
with Src. In contrast,
PMA alone did not potentiate the binding of Src and PKC in the
absence of a translocation event, thereby supporting that not only does
PMA not affect the binding conformation of PKC (14) but
that it also does not directly affect its association with Src. This
latter action is accomplished by PS alone, which alters the
configuration of the substrate- and ATP-binding regions of PKC
(14, 15). These conclusions were further supported by our
data with the PKC open conformation mutants. As was seen in the
presence of PS, the open mutants displayed a higher affinity for
GST-Src than did PKC-WT. The basal interaction between PKC-WT and
GST-Src in the absence of PS may be attributed to the small portion of
PKC-WT that exists in an open conformation.
Contribution of distinct molecular domains.
The next step was to understand what intramolecular domains of PKC
and Src were involved in the interaction of the two molecules. Because
of the fact that previous studies (22) have indicated that
Src interacts with numerous molecules through its SH2 and SH3 domains,
we hypothesized that these domains may regulate its interaction with
PKC. The finding that PKC interacts with both domains (Fig.
1C) is not surprising, because many other molecules that
have been shown to be binding partners of Src family kinases are known
to rely on interaction with these domains (2, 13, 22). The
resultant question was which domain on PKC is responsible for its
interaction with Src. The finding that the regulatory domain of PKC
binds to Src with a much higher affinity than did the catalytic domain
is rather intriguing. Further experiments will be necessary to
understand how this interaction regulates signaling between PKC and Src.
-Src module
formation is consistent with our in vivo findings. We (25) have previously demonstrated that PKC-Src modules are formed in the
rabbit heart and that these modules appear to play a necessary role in
cardioprotection afforded by nitric oxide donors. The essential role of
PKC-Lck modules in PKC-mediated cardioprotection has also been
established by our laboratory in PKC transgenic mice
(22a). Therefore, we wanted to investigate whether
PKC-Src modules were also formed in PKC-cardioprotected mice. The
results gathered in the present study support this notion insofar as
coimmunoprecipitation experiments demonstrate a dramatic increase in
PKC-associated Src in the hearts of cardioprotected mice.
Furthermore, confocal analyses that were performed to asses the
localization of Src revealed that the kinase was distributed across all
subcellular locations. To confirm that these findings were indicative
of Src expression and not due to the nonspecific interactions of a
given antibody, we used multiple antibodies (see METHODS)
directed against distinct epitopes to characterize the distribution of
Src in the heart. The patterns of Src distribution detected by various
antibodies were indistinguishable from each other, supporting that this
kinase is in fact ubiquitously expressed. These confocal analyses
suggest that 1) PKC transgenesis results in increased
distribution of PKC to numerous subcellular compartments,
2) PKC transgenesis induces increased Src expression in
numerous subcellular organelles, and 3) there is a strong
correlation between the subcellular distribution of PKC and Src
after PKC transgenesis. As a result, these findings support the role
of PKC-Src modules as a conserved mechanism of cardiac signaling
across different species.
The importance of the present findings is underscored by the fact that
the PKC-Src module has previously been shown to be an essential
component of the heart's resistance to ischemia after nitric
oxide donors. This point was instrumental in the decision to examine in
the present study the mechanisms that influence module formation
between PKC and Src. Within the cardioprotective PKC signaling
complex, the module containing PKC and Src appears to be an
essential component of protection against cell death. As a result, the
steps taken in this study to understand how this module assembles will
hopefully lead to new strategies to promote assembly in vivo and
thereby enhance the cardioprotective phenotype.
| |
ACKNOWLEDGEMENTS |
|---|
This study was supported in part by National Heart, Lung, and Blood Institute Grants HL-63901 (to P. Ping), HL-65431 (to P. Ping), HL-63760 (to Y. J. Kang), HL-59225 (to Y. J. Kang), HL-66358 (to J. B. Klein), and HL-43721 (to S. D'Souza), by American Heart Association Grant 0110053B (to T. M. Vondriska), and by the University of Louisville Research Foundation, the Department of Veteran's Affairs, and the Jewish Hospital Research Foundation.
| |
FOOTNOTES |
|---|
* These authors contributed equally to this work.
Address for reprint requests and other correspondence: P. Ping, Cardiology Research, Suite 122, Baxter Biomedical Research Bldg., 570 S. Preston St., Louisville, KY 40202 (E-mail: ping{at}ntr.net).
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
10.1152/ajpheart.00830.2001
Received 21 September 2001; accepted in final form 15 November 2001.
| |
REFERENCES |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
1.
Cabodi, S,
Calautti E,
Talora C,
Kuroki T,
Stein PL,
and
Dotto GP.
A PKC-
/Fyn-dependent pathway leading to keratinocyte growth arrest and differentiation.
Mol Cell
6:
1121-1129,
2000[Web of Science][Medline].
2.
Cao, W,
Luttrell LM,
Medvedev AV,
Pierce KL,
Daniel KW,
Dixon TM,
Lefkowitz RJ,
and
Collins S.
Direct binding of activated c-Src to the 
3-adrenergic receptor is required for MAP kinase activation.
J Biol Chem
275:
38131-38134,
2000
3.
Dempsey, EC,
Newton AC,
Mochly-Rosen D,
Fields AP,
Reyland ME,
Insel PA,
and
Messing RO.
Protein kinase C isozymes and the regulation of diverse cell responses.
Am J Physiol Lung Cell Mol Physiol
279:
L429-L438,
2000
4.
Diviani, D,
and
Scott JD.
AKAP signaling complexes at the cytoskeleton.
J Cell Sci
114:
1431-1437,
2001[Abstract].
5.
Dorn, GW, 2nd,
Souroujon MC,
Liron T,
Chen CH,
Gray MO,
Zhou HZ,
Csukai M,
Wu G,
Lorenz JN,
and
Mochly-Rosen D.
Sustained in vivo cardiac protection by a rationally designed peptide that causes epsilon protein kinase C translocation.
Proc Natl Acad Sci USA
96:
12798-12803,
1999
6.
Fryer, RM,
Schultz JE,
Hsu AK,
and
Gross GJ.
Pretreatment with tyrosine kinase inhibitors partially attenuates ischemic preconditioning in rat hearts.
Am J Physiol Heart Circ Physiol
275:
H2009-H2015,
1998
7.
Hattori, R,
Otani H,
Uchiyama T,
Imamura H,
Cui J,
Maulik N,
Cordis GA,
Zhu L,
and
Das DK.
Src tyrosine kinase is the trigger but not the mediator of ischemic preconditioning.
Am J Physiol Heart Circ Physiol
281:
H1066-H1074,
2001
8.
Husi, H,
Ward MA,
Choudhary JS,
Blackstock WP,
and
Grant SG.
Proteomic analysis of NMDA receptor-adhesion protein signaling complexes.
Nat Neurosci
3:
661-669,
2000[Web of Science][Medline].
9.
Kaprielian, RR,
Stevenson S,
Rothery SM,
Cullen MJ,
and
Severs NJ.
Distinct patterns of dystrophin organization in myocyte sarcolemma and transverse tubules of normal and diseased human myocardium.
Circulation
101:
2586-2594,
2000
10.
Kukreja, RC,
Qian YZ,
Okubo S,
and
Flaherty EE.
Role of protein kinase C and 72 kDa heat shock protein in ischemic tolerance following heat stress in the rat heart.
Mol Cell Biochem
195:
123-131,
1999[Web of Science][Medline].
11.
Kwong, KF,
Schuessler RB,
Green KG,
Laing JG,
Beyer EC,
Boineau JP,
and
Saffitz JE.
Differential expression of gap junction proteins in the canine sinus node.
Circ Res
82:
604-612,
1998
12.
Liu, H,
McPherson BC,
and
Yao Z.
Preconditioning attenuates apoptosis and necrosis: role of protein kinase C- and -
isoforms.
Am J Physiol Heart Circ Physiol
281:
H404-H410,
2001
13.
Luttrell, DK,
Lee A,
Lansing TJ,
Crosby RM,
Jung KD,
Willard D,
Luther M,
Rodriquez M,
Berman J,
and
Gilmer TM.
Involvement of pp60c-src with two major signaling pathways in human breast cancer.
Proc Natl Acad Sci USA
91:
83-87,
1994
14.
Newton, AC.
Interaction of proteins with lipid headgroups: lessons from protein kinase C.
Annu Rev Biophys Biomol Struct
22:
1-25,
1993[Web of Science][Medline].
15.
Newton, AC.
Regulation of protein kinase C.
Curr Opin Cell Biol
9:
161-167,
1997[Web of Science][Medline].
16.
Pass, JM,
Zheng Y,
Wead WB,
Zhang J,
Li RC,
Bolli R,
and
Ping P.
PKC activation induces dichotomous cardiac phenotypes and modulates PKC-RACK interactions and RACK expression.
Am J Physiol Heart Circ Physiol
280:
H946-H955,
2001
17.
Phizicky, EM,
and
Fields S.
Protein-protein interactions: methods for detection and analysis.
Microbiol Rev
59:
94-123,
1995
19.
Ping, P,
Zhang J,
Pierce WM, Jr,
and
Bolli R.
Functional proteomic analysis of protein kinase C- signaling complexes in the normal heart and during cardioprotection.
Circ Res
88:
59-62,
2001
20.
Ping, P,
Zhang J,
Qiu Y,
Tang XL,
Manchikalapudi S,
Cao X,
and
Bolli R.
Ischemic preconditioning induces selective translocation of protein kinase C isoforms and in the heart of conscious rabbits without subcellular redistribution of total protein kinase C activity.
Circ Res
81:
404-414,
1997
21.
Ping, P,
Zhang J,
Zheng Y,
Li RC,
Dawn B,
Tang XL,
Takano H,
Balafanova Z,
and
Bolli R.
Demonstration of selective protein kinase C-dependent activation of Src and Lck tyrosine kinases during ischemic preconditioning in conscious rabbits.
Circ Res
85:
542-550,
1999
22.
Sicheri, F,
and
Kuriyan J.
Structures of Src-family kinases.
Curr Opin Struct Biol
7:
777-785,
1997[Web of Science][Medline].
22a.
Song, C,
Zhang J,
Gao J,
Zheng Y,
Cao X,
Wead WB,
Bolli R,
and
Ping P.
Lck tyrosine kinase is a direct substrate of PKC in cardiac myocytes (Abstract).
Circulation
102:
A1021,
2000.
23.
Vahlhaus, C,
Schulz R,
Post H,
Rose J,
and
Heusch G.
Prevention of ischemic preconditioning only by combined inhibition of protein kinase C and protein tyrosine kinase in pigs.
J Mol Cell Cardiol
30:
197-209,
1998[Web of Science][Medline].
24.
Vondriska, TM,
Klein JB,
and
Ping P.
Use of functional proteomics to investigate PKC-mediated cardioprotection: the signaling module hypothesis.
Am J Physiol Heart Circ Physiol
280:
H1434-H1441,
2001
25.
Vondriska, TM,
Zhang J,
Song C,
Tang XL,
Cao X,
Baines CP,
Pass JM,
Wang S,
Bolli R,
and
Ping P.
PKC-Src modules direct signal transduction in NO-induced cardioprotection: complex formation as a means for cardioprotective signaling.
Circ Res
88:
1306-1313,
2001
26.
Ytrehus, K,
Liu Y,
and
Downey JM.
Preconditioning protects ischemic rabbit heart by protein kinase C activation.
Am J Physiol Heart Circ Physiol
266:
H1145-H1152,
1994
This article has been cited by other articles:
|
|
 |
|
  Q. Yu, T. Nguyen, M. Ogbi, R. W. Caldwell, and J. A. Johnson Differential loss of cytochrome-c oxidase subunits in ischemia-reperfusion injury: exacerbation of COI subunit loss by PKC-{varepsilon} inhibition Am J Physiol Heart Circ Physiol, June 1, 2008; 294(6): H2637 - H2645. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Guo, T. Nguyen, M. Ogbi, H. Tawfik, G. Ma, Q. Yu, R. W. Caldwell, and J. A. Johnson Protein kinase C-{varepsilon} coimmunoprecipitates with cytochrome oxidase subunit IV and is associated with improved cytochrome-c oxidase activity and cardioprotection Am J Physiol Heart Circ Physiol, October 1, 2007; 293(4): H2219 - H2230. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. V. Pierre, C. Yang, Z. Yuan, J. Seminerio, C. Mouas, K. D. Garlid, P. Dos-Santos, and Z. Xie Ouabain triggers preconditioning through activation of the Na+,K+-ATPase signaling cascade in rat hearts Cardiovasc Res, February 1, 2007; 73(3): 488 - 496. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Inagaki, E. Churchill, and D. Mochly-Rosen Epsilon protein kinase C as a potential therapeutic target for the ischemic heart Cardiovasc Res, May 1, 2006; 70(2): 222 - 230. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. J. Diaz and G. J. Wilson Studying ischemic preconditioning in isolated cardiomyocyte models Cardiovasc Res, May 1, 2006; 70(2): 286 - 296. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. M. Samarel Costameres, focal adhesions, and cardiomyocyte mechanotransduction Am J Physiol Heart Circ Physiol, December 1, 2005; 289(6): H2291 - H2301. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Zhang, C. P. Baines, C. Zong, E. M. Cardwell, G. Wang, T. M. Vondriska, and P. Ping Functional proteomic analysis of a three-tier PKC{varepsilon}-Akt-eNOS signaling module in cardiac protection Am J Physiol Heart Circ Physiol, February 1, 2005; 288(2): H954 - H961. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. D. Edmondson, T. M. Vondriska, K. J. Biederman, J. Zhang, R. C. Jones, Y. Zheng, D. L. Allen, J. X. Xiu, E. M. Cardwell, M. R. Pisano, et al. Protein Kinase C {epsilon} Signaling Complexes Include Metabolism- and Transcription/Translation-related Proteins: Complimentary Separation Techniques With LC/MS/MS Mol. Cell. Proteomics, June 1, 2002; 1(6): 421 - 433. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| Visit Other APS Journals Online |
