Large scale preparation of calmodulin.

A rapid large scale purification procedure based on three conventional purification steps, ammonium sulfate fractionation, DEAE cellulose and hydroxylapaptite chromotography yields gram amounts of calmodulin. The protein is more than 98% pure by SDS gel electrophoresis and amino acid composition. It is free of contaminating EGTA or EDTA and the omission of heat treatment or denaturing solvents during the preparation avoids possible denaturation of the protein and minimizes partial proteolysis.

[1]  S. Berkowitz Linear multidimensional liquid chromatography in the preparative scale purification of calmodulin from brain extract. , 1987, Analytical biochemistry.

[2]  R. Kretsinger,et al.  Crystal structure of calmodulin. , 1986, Journal of inorganic biochemistry.

[3]  H. Kawasaki,et al.  Limited digestion of calmodulin with trypsin in the presence or absence of various metal ions. , 1986, Journal of biochemistry.

[4]  S. Forsén,et al.  Chapter 4 – Biophysical Studies of Calmodulin , 1986 .

[5]  L. C. Wang,et al.  A simple purification of calmodulin by reversed phase chromatography with hydrophobic polymer 3520. , 1986, Preparative biochemistry.

[6]  R. Kincaid,et al.  Rapid purification of calmodulin and S-100 protein by affinity chromatography with melittin immobilized to sepharose. , 1985, Biochemical and biophysical research communications.

[7]  D. Newton,et al.  Purification and peptide mapping of calmodulin and its chemically modified derivatives by reversed-phase high-performance liquid chromatography. , 1985, Journal of chromatography.

[8]  Charles E. Bugg,et al.  Three-dimensional structure of calmodulin , 1985, Nature.

[9]  W. A. Toscano,et al.  Purification and properties of calmodulin from adrenal cortex. , 1985, Archives of biochemistry and biophysics.

[10]  M. Ikura,et al.  Nuclear magnetic resonance studies on calmodulin: calcium-dependent spectral change of proteolytic fragments , 1984 .

[11]  D. Newton,et al.  Agonist and antagonist properties of calmodulin fragments. , 1984, The Journal of biological chemistry.

[12]  R. J. Williams,et al.  1H NMR studies of calmodulin. Resonance assignments by use of tryptic fragments. , 1984, European journal of biochemistry.

[13]  M. J. Cormier,et al.  Purification of calmodulin by Ca2+-dependent affinity chromatography. , 1983, Methods in enzymology.

[14]  A. Means,et al.  Physiological implications of the presence, distribution, and regulation of calmodulin in eukaryotic cells. , 1982, Physiological reviews.

[15]  J. Cavadore,et al.  Large-scale purification and characterization of calmodulin from ram testis: its metal-ion-dependent conformers. , 1980, Biochimica et biophysica acta.

[16]  K. Seamon Calcium- and magnesium-dependent conformational states of calmodulin as determined by nuclear magnetic resonance. , 1980, Biochemistry.

[17]  W. Y. Cheung,et al.  Calmodulin plays a pivotal role in cellular regulation. , 1980, Science.

[18]  Wai Yiu ed. Cheung,et al.  Calcium and cell function , 1980 .

[19]  C. Klee,et al.  Calcineurin: a calcium- and calmodulin-binding protein of the nervous system. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[20]  C. Klee,et al.  Subunit structure and catalytic properties of bovine brain Ca2+-dependent cyclic nucleotide phosphodiesterase. , 1979, Biochemistry.

[21]  R. Sharma,et al.  Purification of the heat-stable inhibitor protein of the Ca2+-activated cyclic nucleotide phosphodiesterase by affinity chromatography. , 1978, Canadian journal of biochemistry.

[22]  R. Sharma,et al.  Inhibition of Ca2+-activated cyclic nucleotide phosphodiesterase reaction by a heat-stable inhibitor protein from bovine brain. , 1978, The Journal of biological chemistry.

[23]  C. Klee Conformational transition accompanying the binding of Ca2+ to the protein activator of 3',5'-cyclic adenosine monophosphate phosphodiesterase. , 1977, Biochemistry.

[24]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[25]  Ö. Levin [2] Column chromatography of proteins: Calcium phosphate , 1962 .

[26]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.