J. Biochem, 1984, Vol. 95, No. 2 511-519
© 1984 Japanese Biochemical Society
research-article |
Detection of Calcium Binding Proteins by 45Ca Autoradiography on Nitrocellulose Membrane after Sodium Dodecyl Sulfate Gel Electrophoresis1
Department of Pharmacology, Faculty of Medicine, The University of Tokyo Hongo, Bunkyo-ku, Tokyo 113
A new simple method of detecting calcium binding proteins in a protein mixture is described. A sample which might include calcium binding proteins was subjected to SDS-polyacrylamide gel electrophoresis and then electrophoretically transferred to a nitrocellulose membrane. The membrane was then incubated with 45Ca to detect calcium binding proteins as radioactive bands by autoradiography.
Purified troponin-C, calmodulin, myosin DTNB light chain, and parvalbumin were clearly identified by this method. In the whole homogenate of chicken skeletal muscle, myosin DTNB light chain, troponin-C, and 55K calcium binding protein were found to be radioactive. In the frog skeletal muscle, small molecular weight proteins of approximately 13–15K and 70K protein appeared to be the calcium binding proteins. In the case of the carp skeletal muscle, small molecular weight proteins including parvalbumin and two proteins of about 80K seemed to bind calcium ion. Two high molecular weight calcium binding proteins were present in the scallop striated muscle.
The procedure described can be completed within 24 h and can detect as little as 2 µg of calcium binding protein in the starting sample. Under appropriate conditions it was possible to detect only high affinity calcium binding proteins.
1 This work was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan, the Muscular Dystrophy Association of America, and the Tatrochemical Research Foundation.
2 Present address: Laboratory of Neurochemistry, National lnstitute for Physiological Sciences, Myodaiji, Oka-zaki, Aichi 444.
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T. Nguyen and T. C. Sudhof Binding Properties of Neuroligin 1 and Neurexin 1beta Reveal Function as Heterophilic Cell Adhesion Molecules J. Biol. Chem., October 10, 1997; 272(41): 26032 - 26039. [Abstract] [Full Text] [PDF]
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H. He, M. Lam, T. S. McCormick, and C. W. Distelhorst Maintenance of Calcium Homeostasis in the Endoplasmic Reticulum by Bcl-2 J. Cell Biol., September 22, 1997; 138(6): 1219 - 1228. [Abstract] [Full Text] [PDF]
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H. Tran, W. J. VanDusen, and W. S. Argraves The Self-association and Fibronectin-binding Sites of Fibulin-1 Map to Calcium-binding Epidermal Growth Factor-like Domains J. Biol. Chem., September 5, 1997; 272(36): 22600 - 22606. [Abstract] [Full Text] [PDF]
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F. E. Weber, G. Minestrini, J. H. Dyer, M. Werder, D. Boffelli, S. Compassi, E. Wehrli, R. M. Thomas, G. Schulthess, and H. Hauser Molecular cloning of a peroxisomal Ca2+-dependent member of the mitochondrial carrier superfamily PNAS, August 5, 1997; 94(16): 8509 - 8514. [Abstract] [Full Text] [PDF]
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D. Yabe, T. Nakamura, N. Kanazawa, K. Tashiro, and T. Honjo Calumenin, a Ca2+-binding Protein Retained in the Endoplasmic Reticulum with a Novel Carboxyl-terminal Sequence, HDEF J. Biol. Chem., July 18, 1997; 272(29): 18232 - 18239. [Abstract] [Full Text] [PDF]
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P. Falson, T. Menguy, F. Corre, L. Bouneau, A. G. de Gracia, S. Soulie, F. Centeno, J. V. Moller, P. Champeil, and M. le Maire The Cytoplasmic Loop between Putative Transmembrane Segments 6 and 7 in Sarcoplasmic Reticulum Ca2+-ATPase Binds Ca2+ and Is Functionally Important J. Biol. Chem., July 11, 1997; 272(28): 17258 - 17262. [Abstract] [Full Text] [PDF]
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T. D. Spurway, C. Morland, A. Cooper, I. Sumner, G. P. Hazlewood, A. G. O'Donnell, R. W. Pickersgill, and H. J. Gilbert Calcium Protects a Mesophilic Xylanase from Proteinase Inactivation and Thermal Unfolding J. Biol. Chem., July 11, 1997; 272(28): 17523 - 17530. [Abstract] [Full Text] [PDF]
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G. Siegenthaler, K. Roulin, D. Chatellard-Gruaz, R. Hotz, J. H. Saurat, U. Hellman, and G. Hagens A Heterocomplex Formed by the Calcium-binding Proteins MRP8 (S100A8) and MRP14 (S100A9) Binds Unsaturated Fatty Acids with High Affinity J. Biol. Chem., April 4, 1997; 272(14): 9371 - 9377. [Abstract] [Full Text] [PDF]
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K. Scholich, A. J. Barbier, J. B. Mullenix, and T. B. Patel Characterization of soluble forms of nonchimeric type V adenylyl cyclases PNAS, April 1, 1997; 94(7): 2915 - 2920. [Abstract] [Full Text] [PDF]
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U. P. Naik, P. M. Patel, and L. V. Parise Identification of a Novel Calcium-binding Protein That Interacts with the Integrin alpha IIb Cytoplasmic Domain J. Biol. Chem., February 21, 1997; 272(8): 4651 - 4654. [Abstract] [Full Text] [PDF]
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M. S. Airaksinen, J. Eilers, O. Garaschuk, H. Thoenen, A. Konnerth, and M. Meyer Ataxia and altered dendritic calcium signaling in mice carrying a targeted null mutation of the calbindin D28k gene PNAS, February 18, 1997; 94(4): 1488 - 1493. [Abstract] [Full Text] [PDF]
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M. A.C. Silva-Neto, G. C. Atella, E. Fialho, M. C. Paes, R. B. Zingali, J. H. Petretski, E. W. Alves, and H. Masuda Isolation of a Calcium-binding Phosphoprotein from the Oocytes and Hemolymph of the Blood-sucking Insect Rhodnius prolixus J. Biol. Chem., November 22, 1996; 271(47): 30227 - 30232. [Abstract] [Full Text] [PDF]
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I. Sienaert, H. De Smedt, J. B. Parys, L. Missiaen, S. Vanlingen, H. Sipma, and R. Casteels Characterization of a Cytosolic and a Luminal Ca2+ Binding Site in the Type I Inositol 1,4,5-Trisphosphate Receptor J. Biol. Chem., October 25, 1996; 271(43): 27005 - 27012. [Abstract] [Full Text] [PDF]
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F. Zorzato, P. Menegazzi, S. Treves, and M. Ronjat Role of Malignant Hyperthermia Domain in the Regulation of Ca2+ Release Channel (Ryanodine Receptor) of Skeletal Muscle Sarcoplasmic Reticulum J. Biol. Chem., September 13, 1996; 271(37): 22759 - 22763. [Abstract] [Full Text] [PDF]
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Z. Yang, M. J. deVeer, E. E. Gardiner, R. J. Devenish, C. J. Handley, J. R. Underwood, and H. C. Robinson Rabbit Polymorphonuclear Neutrophils Form 35S-Labeled S-Sulfo-calgranulin C When Incubated with Inorganic [35S]Sulfate J. Biol. Chem., August 16, 1996; 271(33): 19802 - 19809. [Abstract] [Full Text] [PDF]
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E. F.S. Wong, S. K. Brar, H. Sesaki, C. Yang, and C.-H. Siu Molecular Cloning and Characterization of DdCAD-1, a Ca2+-dependent Cell-Cell Adhesion Molecule, in Dictyostelium discoideum J. Biol. Chem., July 5, 1996; 271(27): 16399 - 16408. [Abstract] [Full Text] [PDF]
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G. Patstone and P. Maher Copper and Calcium Binding Motifs in the Extracellular Domains of Fibroblast Growth Factor Receptors J. Biol. Chem., February 16, 1996; 271(7): 3343 - 3346. [Abstract] [Full Text] [PDF]
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C. Realini and M. Rechsteiner A Proteasome Activator Subunit Binds Calcium J. Biol. Chem., December 15, 1995; 270(50): 29664 - 29667. [Abstract] [Full Text] [PDF]
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T. Ohta, M. Kobayashi, and S. Hirose Cloning of a cDNA for DNA Supercoiling Factor Reveals a Distinctive Ca[IMAGE]-binding Protein J. Biol. Chem., June 30, 1995; 270(26): 15571 - 15575. [Abstract] [Full Text] [PDF]
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M. Wendel, Y. Sommarin, T. Bergman, and D. Heinegård Isolation, Characterization, and Primary Structure of a Calcium-binding 63-kDa Bone Protein J. Biol. Chem., March 17, 1995; 270(11): 6125 - 6133. [Abstract] [Full Text] [PDF]
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O. M. Andersen, L. L. Christensen, P. A. Christensen, E. S. Sorensen, C. Jacobsen, S. K. Moestrup, M. Etzerodt, and H. C. Thogersen Identification of the Minimal Functional Unit in the Low Density Lipoprotein Receptor-related Protein for Binding the Receptor-associated Protein (RAP). A CONSERVED ACIDIC RESIDUE IN THE COMPLEMENT-TYPE REPEATS IS IMPORTANT FOR RECOGNITION OF RAP J. Biol. Chem., July 7, 2000; 275(28): 21017 - 21024. [Abstract] [Full Text] [PDF]
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N. Miwa, T. Uebi, and S. Kawamura Characterization of p26olf, a Novel Calcium-binding Protein in the Frog Olfactory Epithelium J. Biol. Chem., August 25, 2000; 275(35): 27245 - 27249. [Abstract] [Full Text] [PDF]
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F. Marin, P. Corstjens, B. de Gaulejac, E. de Vrind-De Jong, and P. Westbroek Mucins and Molluscan Calcification. MOLECULAR CHARACTERIZATION OF MUCOPERLIN, A NOVEL MUCIN-LIKE PROTEIN FROM THE NACREOUS SHELL LAYER OF THE FAN MUSSEL PINNA NOBILIS (BIVALVIA, PTERIOMORPHIA) J. Biol. Chem., June 30, 2000; 275(27): 20667 - 20675. [Abstract] [Full Text] [PDF]
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Y. Jiang, W. Qian, J. W. Hawes, and J. P. Walsh A Domain with Homology to Neuronal Calcium Sensors Is Required for Calcium-dependent Activation of Diacylglycerol Kinase alpha J. Biol. Chem., October 27, 2000; 275(44): 34092 - 34099. [Abstract] [Full Text] [PDF]
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N. Taniguchi, H. Taniura, M. Niinobe, C. Takayama, K. Tominaga-Yoshino, A. Ogura, and K. Yoshikawa The Postmitotic Growth Suppressor Necdin Interacts with a Calcium-binding Protein (NEFA) in Neuronal Cytoplasm J. Biol. Chem., October 6, 2000; 275(41): 31674 - 31681. [Abstract] [Full Text] [PDF]
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S. Treves, G. Feriotto, L. Moccagatta, R. Gambari, and F. Zorzato Molecular Cloning, Expression, Functional Characterization, Chromosomal Localization, and Gene Structure of Junctate, a Novel Integral Calcium Binding Protein of Sarco(endo)plasmic Reticulum Membrane J. Biol. Chem., December 8, 2000; 275(50): 39555 - 39568. [Abstract] [Full Text] [PDF]
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Y. Bourne, J. Dannenberg, V. Pollmann, P. Marchot, and O. Pongs Immunocytochemical Localization and Crystal Structure of Human Frequenin (Neuronal Calcium Sensor 1) J. Biol. Chem., April 6, 2001; 276(15): 11949 - 11955. [Abstract] [Full Text] [PDF]
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S. Bian, I. Favre, and E. Moczydlowski Ca2+-binding activity of a COOH-terminal fragment of the Drosophila BK channel involved in Ca2+-dependent activation PNAS, April 10, 2001; 98(8): 4776 - 4781. [Abstract] [Full Text] [PDF]
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C. Xi, E. Schoeters, J. Vanderleyden, and J. Michiels Symbiosis-specific expression of Rhizobium etli casA encoding a secreted calmodulin-related protein PNAS, September 26, 2000; 97(20): 11114 - 11119. [Abstract] [Full Text] [PDF]
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