Smooth muscle α-actinin binds tightly to fesselin and attenuates its activity toward actin polymerization
- 51 Downloads
- 8 Citations
Abstract
Fesselin is an actin binding protein from smooth muscle that nucleates actin polymerization in a Ca++-calmodulin dependent manner, bundles actin and inhibits the actin-activated ATPase activity of myosin S1. We now report that fesselin binds to smooth muscle α-actinin. Binding was measured by blot overlay, affinity chromatography and sedimentation methods. Binding was moderate with an association constant of 1–4×107 M−1 assuming a 1:1 association of fesselin with α-actinin. Fesselin binds to the central spectrin domain repeat region of α-actinin but not to the CH1–CH2 domain. Fesselin accelerates the polymerization of actin. This activity of fesselin was attenuated by α-actinin. These observations support the role of fesselin in organizing the cytoskeleton.
Keywords
Actin Polymerization Bundle Actin Pyrenyl Calponin Homology Domain Spectrin RepeatNotes
Acknowledgements
The authors thank Dr. Roberto Dominguez for his gift of the CH1–CH2 domain of α-actinin and Ms. Natalie Lonergan and Ms. Tamatha Baxley for technical assistance. This work was supported by grant AR35216 from the National Institutes of Health to JMC.
References
- Asanuma K, Kim K, Oh J, Giardino L, Chabanis S, Faul C, Reiser J, Mundel P, (2005). Synaptopodin regulates the actin-bundling activity of α-actinin in an isoform-specific manner J Clin Invest 115:1188–1198PubMedGoogle Scholar
- Baron MD, Davison MD, Jones P, Critchley DR, (1987). The sequence of chick alpha actinin reveals homologies to spectrin and calmodulin J Biol Chem 262:17623–17629PubMedGoogle Scholar
- Beall B, Chalovich JM, (2001). Fesselin a synaptopodin-like protein, stimulates actin nucleation and polymerization Biochemistry 40:14252–14259PubMedCrossRefGoogle Scholar
- Blanchard A, Ohanian V, Critchley D, (1989). The structure and function of alpha-actinin J Muscle Res Cell Motil 10:280–289PubMedCrossRefGoogle Scholar
- Brenner SL, Korn ED, (1983). On the mechanism of actin monomer–polymer subunit exchange at steady state J Biol Chem 258:5013–5020PubMedGoogle Scholar
- Critchley DR, Flood G, (1999). α-actinins In: Kreis T, Vale R, (eds). Guidebook to the Cytoskeletal and Motor Proteins (2nd ed). Oxford University Press New York 24–27Google Scholar
- Eisenberg E, Kielley WW, (1972). Evidence for a refractory state of heavy meromyosin and subfragment-1 unable to bind to actin in the presence of ATP Cold Spring Harbor Symp Quant Biol 37:145–152Google Scholar
- Feramisco JR, Burridge K, (1980). A rapid purification of a-actinin, filamin, and a 130,000-dalton protein from smooth muscle J Biol Chem 255:1194–1199PubMedGoogle Scholar
- Fraley TS, Tran TC, Corgan AM, Nash CA, Hao J, Critchley DR, Greenwood JA, (2003). Phosphoinositide binding inhibits α-actinin bundling activity J Biol Chem 278:24039–24045PubMedCrossRefGoogle Scholar
- Imamura M, Endo T, Kuroda M, Tanaka T, Masaki T (1988). Substructure and higher structure of chicken smooth muscle alpha-actinin molecule J Biol Chem 263:7800–7805PubMedGoogle Scholar
- Kolakowski J, Wrzosek A, Dabrowska R, (2004). Fesselin is a target protein for calmodulin in a calcium-dependent manner Biochem Biophys Res Commun 323:1251–1256PubMedCrossRefGoogle Scholar
- Kouyama T, Mihashi K, (1981). Fluorimetry study of N-(1-pyrenyl)iodoacetamide-labeled F-actin: local structural change of actin protomer both on polymerization and on binding of heavy meromyosin Eur J Biochem 114:33–38PubMedGoogle Scholar
- Kremerskothen J, Plaas C, Kindler S, Frotscher M, Barnekow A, (2005). Synaptopodin, a molecule involved in the formation of the dendritic spine apparatus, is a dual actin/α-actinin binding protein J Neurochem 92:597–606PubMedCrossRefGoogle Scholar
- Kuroda M, Kohira Y, Sasaki M, (1994). Conformational change of skeletal muscle alpha-actinin induced by salt Biochim Biophys Acta 1205:97–104PubMedGoogle Scholar
- Leinweber, B. (1997) Ph.D. thesis, Maintaining the localization of actin in smooth muscle: α-actinin, filamin, calponin and SM85/95. Brody School of Medicine at East Carolina University, Greenville, NCGoogle Scholar
- Leinweber BD, Fredricksen RS, Hoffman DR, Chalovich JM, (1999). Fesselin: a novel synaptopodin-like actin binding protein from muscle tissue J Muscle Res Cell Motil 20:539–545PubMedCrossRefGoogle Scholar
- Leinweber B, Tang JX, Stafford WF, Chalovich JM, (1999). Calponin interaction with α-actinin–actin: Evidence for a structural role for calponin Biophys J 77:3208–3217PubMedCrossRefGoogle Scholar
- Mundel P, Heid HW, Mundel TM, Krüger M, Reiser J, Kriz W, (1997). Synaptopodin: An actin-associated protein in telencephalic dendrites and renal podocytes J Cell Biol 139:193–204PubMedCrossRefGoogle Scholar
- Otey CA, Carpen O, (2004). α-Actinin revisited: a fresh look at an old player Cell Motil Cytoskel 58:104–111CrossRefGoogle Scholar
- Patrie KM, Drescher AJ, Welihinda A, Mundel P, Margolis B, (2002). Interaction of two actin-binding proteins, synaptopodin and α-actinin-4, with the tight junction protein MAGI-1, J Biol Chem 277:30183–30190PubMedCrossRefGoogle Scholar
- Pollard TD, Cooper JA (1982). Methods to characterize actin filament networks Method Enzymol 85:211–233Google Scholar
- Schroeter M, Chalovich JM, (2004). Ca2+-calmodulin regulates fesselin-induced actin polymerization Biochemistry 43:13875–13882PubMedCrossRefGoogle Scholar
- Schroeter MM and Chalovich JM (2005) Fesselin binds to actin and myosin and inhibits actin activated ATPase activity, J Muscle Res Cell Motil 26:183–189PubMedCrossRefGoogle Scholar
- Spudich JA, Watt S, (1971). The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin–troponin complex with actin and the proteolytic fragments of myosin J Biol Chem 246:4866–4871PubMedGoogle Scholar
- Weins A, Schwarz K, Faul C, Barisoni L, Linke WA, Mundel P, (2001). Differentiation- and stress-dependent nuclear cytoplasmic redistribution of myopodin, a novel actin-bundling protein J Cell Biol 155:393–404PubMedCrossRefGoogle Scholar