Abstract
2-(5'-Phenylpentyl)fumaric acid was shown to be a competitive inhibitor (Ki 0.5 mM) of pig heart fumarase. After nitration of the aromatic ring, reduction to the amine and diazotization, the acid was attached via azo linkages to a Sepharose 4B-tyramine matrix. The resulting adsorbent was used for the affinity chromatography of crude fumarase, purifications of approx. 20-fold being obtained by specific elution with 0.01 M-citrate.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Beeckmans S., Kanarek L. A new purification procedure for fumarase based of affinity chromatography. Isolation and characterization of pig-liver fumarase. Eur J Biochem. 1977 Sep;78(2):437–444. doi: 10.1111/j.1432-1033.1977.tb11756.x. [DOI] [PubMed] [Google Scholar]
- Cuatrecasas P. Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J Biol Chem. 1970 Jun;245(12):3059–3065. [PubMed] [Google Scholar]
- DIXON M. The determination of enzyme inhibitor constants. Biochem J. 1953 Aug;55(1):170–171. doi: 10.1042/bj0550170. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lin Y. C., Scott C. F., Cohen L. H. Multiple forms of fumarase of pig heart. Arch Biochem Biophys. 1971 Jun;144(2):741–748. doi: 10.1016/0003-9861(71)90382-1. [DOI] [PubMed] [Google Scholar]
- MASSEY V. The crystallization of fumarase. Biochem J. 1952 Jul;51(4):490–494. doi: 10.1042/bj0510490. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rogers G. A., Shaltiel N., Boyer P. D. Facile alkylation of methionine by benzyl bromide and demonstration of fumarase inactivation accompanied by alkylation of a methionine residue. J Biol Chem. 1976 Sep 25;251(18):5711–5717. [PubMed] [Google Scholar]
- Teipel J. W., Hass G. M., Hill R. L. The substrate specificity of fumarase. J Biol Chem. 1968 Nov 10;243(21):5684–5694. [PubMed] [Google Scholar]