Publications

• Structure determination and refinement at 2.44 A resolution of argininosuccinate lyase from Escherichia coli.

  Bhaumik P., Koski M.K., Bergmann U., Wierenga R.K.

  Escherichia coli argininosuccinate lyase has been crystallized from a highly concentrated sample of purified recombinant alpha-methylacyl-CoA racemase, in which it occurred as a minor impurity. The structure has been solved using molecular replacement at 2.44 A resolution. The enzyme is tetrameric ... >> More

  Escherichia coli argininosuccinate lyase has been crystallized from a highly concentrated sample of purified recombinant alpha-methylacyl-CoA racemase, in which it occurred as a minor impurity. The structure has been solved using molecular replacement at 2.44 A resolution. The enzyme is tetrameric, but in this crystal form there is a dimer in the asymmetric unit. The tetramer has four active sites; each active site is constructed from loops of three different subunits. One of these catalytic loops, near residues Ser277 and Ser278, was disordered in previous structures of active lyases, but is very well ordered in this structure in one of the subunits owing to the presence of two phosphate ions in the active-site cavity. The positions of these phosphate ions indicate a plausible mode of binding of the succinate moiety of the substrate in the competent catalytic complex. << Less

  Acta Crystallogr D Biol Crystallogr 60:1964-1970(2004) [PubMed] [EuropePMC]

• Enzymic reaction between arginine and fumarate in plant and animal tissues.

  DAVISON D.C., ELLIOTT W.H.

  Nature 169:313-314(1952) [PubMed] [EuropePMC]

• Characterization of argininosuccinate lyase (EC 4.3.2.1) from Chlamydomonas reinhardtii.

  Farrell K., Overton S.

  We have isolated and characterized argininosuccinate lyase (ASL; EC 4.3.2.1) from the photosynthetic green alga, Chlamydomonas reinhardtii. The general properties of Chlamydomonas ASL are very similar to those described previously for ASLs from phylogenetically diverse organisms. The algal ASL has ... >> More

  We have isolated and characterized argininosuccinate lyase (ASL; EC 4.3.2.1) from the photosynthetic green alga, Chlamydomonas reinhardtii. The general properties of Chlamydomonas ASL are very similar to those described previously for ASLs from phylogenetically diverse organisms. The algal ASL has a native Mr, determined by gel-filtration chromatography, of 218,000 +/-25,000, and a pI of 5.4-5.6. The Km for argininosuccinate at 37 degrees C and pH 7.5 is 0.26 mM. The subunit Mr of Chlamydomonas ASL is approx. 50,000, determined by SDS/polyacrylamide-gel electrophoresis, in contrast with a previously reported value of 39,000. Rabbit antisera prepared against the Mr-50,000 protein completely abolished ASL activity in vitro. In contrast, serum prepared against the Mr-39,000 protein was ineffective in inhibiting ASL activity. Despite the general similarity of the physical properties of Chlamydomonas ASL and those of other ASLs, antiserum raised against the algal ASL did not cross-react with ASL preparations from Escherichia coli, Saccharomyces cerevisiae or bovine liver. << Less

  Biochem J 242:261-266(1987) [PubMed] [EuropePMC]

• Argininosuccinate lyase: purification and characterization from human liver.

  O'Brien W.E., Barr R.H.

  Argininosuccinate lyase has been purified to near homogeneity and partially characterized from extracts of human liver. The purified enzyme had a specific activity of 10.3 mumol min-1 mg-1 in the forward, argininosuccinate cleaving, reaction and 8.0 mumol min-1 mg-1 in the reverse reaction. On the ... >> More

  Argininosuccinate lyase has been purified to near homogeneity and partially characterized from extracts of human liver. The purified enzyme had a specific activity of 10.3 mumol min-1 mg-1 in the forward, argininosuccinate cleaving, reaction and 8.0 mumol min-1 mg-1 in the reverse reaction. On the basis of electrophoretic mobility in sodium dodecyl sulfate containing polyacrylamide gels, the protein had a minimum molecular weight of 49 000. Sedimentation equilibrium centrifugation revealed a molecular weight of 187 000. On the basis of these data, the enzyme appears to be a tetramer composed of subunits of identical molecular weight. The Km values were about 0.20 mM for argininosuccinate, 5.3 mM for fumarte, and 3.0 mM for arginine. The enzyme exhibited normal Michaelis-Menten kinetics, and guanosine 5'-triphosphate (GTP) had no affect on the activity of the enzyme. With the exception of its kinetic properties, the enzyme is very similar to the beef liver enzyme. Antibodies were prepared in rabbits and were specific for the human protein, reacting only slightly with the beef liver enzyme and not at all with the rat liver enzyme. The antibodies reacted identically with purified enzyme and enzyme in extracts of human skin fibroblasts. Immunoadsorption of crude human liver extracts followed by analysis of the immunoprecipitates by sodium dodecyl sulfate gel electrophoresis showed only one protein band which corresponded in mobility to purified argininosuccinate lyase, demonstrating that the antibodies were specific for argininosuccinate lyase. << Less

  Biochemistry 20:2056-2060(1981) [PubMed] [EuropePMC]