Abstract
1-Aminocyclopropane-1-carboxylic acid (ACC) N-malonyltransferase catalyzes the transfer of the malonyl group from malonyl coenzyme A to ACC to form malonyl ACC. Using partially purified ACC N-malonyltransferase from the hypocotyls of mung bean (Vigna radiata) seedlings, we produced two mouse monoclonal antibodies (1H5 and 2G3) to this enzyme. These antibodies bind to sites other than the active site of the enzyme because monoclonal antibody-bound ACC N-malonyltransferase still exhibits full catalytic activity. A monoclonal antibody column was constructed using 1H5 and protein G Sepharose. The ACC N-malonyltransferase purified from this monoclonal antibody column has a molecular mass of 40 kD, which is different from that reported previously. The enzyme has a higher electrophoretic mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the absence of the reducing agent dithiothreitol. The optimum temperature of this 40-kD ACC N-malonyltransferase is 45 degrees C and the apparent Kms for ACC and malonyl coenzyme A are 66.7 and 40 microns, respectively.
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- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Galfrè G., Milstein C. Preparation of monoclonal antibodies: strategies and procedures. Methods Enzymol. 1981;73(Pt B):3–46. doi: 10.1016/0076-6879(81)73054-4. [DOI] [PubMed] [Google Scholar]
- Guo L., Arteca R. N., Phillips A. T., Liu Y. Purification and characterization of 1-aminocyclopropane-1-carboxylate N-malonyltransferase from etiolated mung bean hypocotyls. Plant Physiol. 1992 Dec;100(4):2041–2045. doi: 10.1104/pp.100.4.2041. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Guo L., Phillips A. T., Arteca R. N. Amino acid N-malonyltransferases from mung beans. Action on 1-aminocyclopropane-1-carboxylic acid and D-phenylalanine. J Biol Chem. 1993 Dec 5;268(34):25389–25394. [PubMed] [Google Scholar]
- Hoffman N. E., Yang S. F., McKeon T. Identification of 1-(malonylamino) cyclopropane-1-carboxylic acid as a major conjugate of 1-aminocyclopropane-1-carboxylic acid, an ethylene precursor in higher plants. Biochem Biophys Res Commun. 1982 Jan 29;104(2):765–770. doi: 10.1016/0006-291x(82)90703-3. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Martin M. N., Saftner R. A. Purification and Characterization of 1-Aminocyclopropane-1-Carboxylic Acid N-Malonyltransferase from Tomato Fruit. Plant Physiol. 1995 Jul;108(3):1241–1249. doi: 10.1104/pp.108.3.1241. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matern U., Feser C., Heller W. N-malonyltransferases from peanut. Arch Biochem Biophys. 1984 Nov 15;235(1):218–227. doi: 10.1016/0003-9861(84)90271-6. [DOI] [PubMed] [Google Scholar]