Activation of M-calpain (calpain II) by Epidermal Growth Factor is Limited by Protein Kinase A Phosphorylation of M-calpain

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 2002 Mar 23

PMID 11909964

Citations 68

Authors

Hidenori Shiraha

Angela Glading

Jeffrey Chou

Zongchao Jia

Alan Wells

Affiliations

Soon will be listed here.

Abstract

We have shown previously that the ELR-negative CXC chemokines interferon-inducible protein 10, monokine induced by gamma interferon, and platelet factor 4 inhibit epidermal growth factor (EGF)-induced m-calpain activation and thereby EGF-induced fibroblast cell motility (H. Shiraha, A. Glading, K. Gupta, and A. Wells, J. Cell Biol. 146:243-253, 1999). However, how this cross attenuation could be accomplished remained unknown since the molecular basis of physiological m-calpain regulation is unknown. As the initial operative attenuation signal from the CXCR3 receptor was cyclic AMP (cAMP), we verified that this second messenger blocked EGF-induced motility of fibroblasts (55% +/- 4.5% inhibition) by preventing rear release during active locomotion. EGF-induced calpain activation was inhibited by cAMP activation of protein kinase A (PKA), as the PKA inhibitors H-89 and Rp-8Br-cAMPS abrogated cAMP inhibition of both motility and calpain activation. We hypothesized that PKA might negatively modulate m-calpain in an unexpected manner by directly phosphorylating m-calpain. A mutant human large subunit of m-calpain was genetically engineered to negate a putative PKA consensus sequence in the regulatory domain III (ST369/370AA) and was expressed in NR6WT mouse fibroblasts to represent about 30% of total m-calpain in these cells. This construct was not phosphorylated by PKA in vitro while a wild-type construct was, providing proof of the principle that m-calpain can be directly phosphorylated by PKA at this site. cAMP suppressed EGF-induced calpain activity of cells overexpressing a control wild-type human m-calpain (83% +/- 3.7% inhibition) but only marginally suppressed that of cells expressing the PKA-resistant mutant human m-calpain (25% +/- 5.5% inhibition). The EGF-induced motility of the cells expressing the PKA-resistant mutant also was not inhibited by cAMP. Structural modeling revealed that new constraints resulting from phosphorylation at serine 369 would restrict domain movement and help "freeze" m-calpain in an inactive state. These data point to a novel mechanism of negative control of calpain activation, direct phosphorylation by PKA.

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References

Jia Z, Petrounevitch V, Wong A, Moldoveanu T, Davies P, Elce J . Mutations in calpain 3 associated with limb girdle muscular dystrophy: analysis by molecular modeling and by mutation in m-calpain. Biophys J. 2001; 80(6):2590-6. PMC: 1301447. DOI: 10.1016/S0006-3495(01)76229-7. View

Imajoh S, Aoki K, Ohno S, Emori Y, Kawasaki H, Sugihara H . Molecular cloning of the cDNA for the large subunit of the high-Ca2+-requiring form of human Ca2+-activated neutral protease. Biochemistry. 1988; 27(21):8122-8. DOI: 10.1021/bi00421a022. View

Hosfield C, Moldoveanu T, Davies P, Elce J, Jia Z . Calpain mutants with increased Ca2+ sensitivity and implications for the role of the C(2)-like domain. J Biol Chem. 2000; 276(10):7404-7. DOI: 10.1074/jbc.M007352200. View

Lane R, Allan D, Mellgren R . A comparison of the intracellular distribution of mu-calpain, m-calpain, and calpastatin in proliferating human A431 cells. Exp Cell Res. 1992; 203(1):5-16. DOI: 10.1016/0014-4827(92)90033-5. View

Goebeler M, Yoshimura T, Toksoy A, Ritter U, Brocker E, Gillitzer R . The chemokine repertoire of human dermal microvascular endothelial cells and its regulation by inflammatory cytokines. J Invest Dermatol. 1997; 108(4):445-51. DOI: 10.1111/1523-1747.ep12289711. View

Engelhardt E, Toksoy A, Goebeler M, Debus S, Brocker E, Gillitzer R . Chemokines IL-8, GROalpha, MCP-1, IP-10, and Mig are sequentially and differentially expressed during phase-specific infiltration of leukocyte subsets in human wound healing. Am J Pathol. 1998; 153(6):1849-60. PMC: 1866330. DOI: 10.1016/s0002-9440(10)65699-4. View

JOHNSON G, Foley V . Calpain-mediated proteolysis of microtubule-associated protein 2 (MAP-2) is inhibited by phosphorylation by cAMP-dependent protein kinase, but not by Ca2+/calmodulin-dependent protein kinase II. J Neurosci Res. 1993; 34(6):642-7. DOI: 10.1002/jnr.490340607. View

Adachi Y, Kobayashi N, Murachi T, Hatanaka M . Ca2+-dependent cysteine proteinase, calpains I and II are not phosphorylated in vivo. Biochem Biophys Res Commun. 1986; 136(3):1090-6. DOI: 10.1016/0006-291x(86)90445-6. View

Ware M, Wells A, Lauffenburger D . Epidermal growth factor alters fibroblast migration speed and directional persistence reciprocally and in a matrix-dependent manner. J Cell Sci. 1998; 111 ( Pt 16):2423-32. DOI: 10.1242/jcs.111.16.2423. View

10.

Hirose K, Kadowaki S, Tanabe M, Takeshima H, Iino M . Spatiotemporal dynamics of inositol 1,4,5-trisphosphate that underlies complex Ca2+ mobilization patterns. Science. 1999; 284(5419):1527-30. DOI: 10.1126/science.284.5419.1527. View

11.

Sorimachi H, Ishiura S, Suzuki K . Structure and physiological function of calpains. Biochem J. 1998; 328 ( Pt 3):721-32. PMC: 1218978. DOI: 10.1042/bj3280721. View

12.

Cole K, Strick C, Paradis T, Ogborne K, Loetscher M, Gladue R . Interferon-inducible T cell alpha chemoattractant (I-TAC): a novel non-ELR CXC chemokine with potent activity on activated T cells through selective high affinity binding to CXCR3. J Exp Med. 1998; 187(12):2009-21. PMC: 2212354. DOI: 10.1084/jem.187.12.2009. View

13.

McClelland P, Adam L, Hathaway D . Identification of a latent Ca2+/calmodulin dependent protein kinase II phosphorylation site in vascular calpain II. J Biochem. 1994; 115(1):41-6. DOI: 10.1093/oxfordjournals.jbchem.a124302. View

14.

Cook S, McCormick F . Inhibition by cAMP of Ras-dependent activation of Raf. Science. 1993; 262(5136):1069-72. DOI: 10.1126/science.7694367. View

15.

Strobl S, Braun M, Huber R, Masumoto H, Nakagawa K, Irie A . The crystal structure of calcium-free human m-calpain suggests an electrostatic switch mechanism for activation by calcium. Proc Natl Acad Sci U S A. 2000; 97(2):588-92. PMC: 15374. DOI: 10.1073/pnas.97.2.588. View

16.

Rosser B, Powers S, Gores G . Calpain activity increases in hepatocytes following addition of ATP. Demonstration by a novel fluorescent approach. J Biol Chem. 1993; 268(31):23593-600. View

17.

Carafoli E, Molinari M . Calpain: a protease in search of a function?. Biochem Biophys Res Commun. 1998; 247(2):193-203. DOI: 10.1006/bbrc.1998.8378. View

18.

Froehlich J, Rachmeler M . Effect of adenosine 3'-5'-cyclic monophosphate on cell proliferation. J Cell Biol. 1972; 55(1):19-31. PMC: 2108750. DOI: 10.1083/jcb.55.1.19. View

19.

Du X, Saido T, Tsubuki S, Indig F, WILLIAMS M, Ginsberg M . Calpain cleavage of the cytoplasmic domain of the integrin beta 3 subunit. J Biol Chem. 1995; 270(44):26146-51. DOI: 10.1074/jbc.270.44.26146. View

20.

JOHNSON G, Jope R, Binder L . Proteolysis of tau by calpain. Biochem Biophys Res Commun. 1989; 163(3):1505-11. DOI: 10.1016/0006-291x(89)91150-9. View