Literature DB >> 24598918

Purification, crystallization and preliminary crystallographic analysis of the catalytic core of cystathionine β-synthase from Saccharomyces cerevisiae.

June Ereño-Orbea1, Tomas Majtan2, Iker Oyenarte1, Jan P Kraus2, Luis Alfonso Martínez-Cruz1.   

Abstract

Cystathionine β-synthase (CBS; EC 4.2.1.22) catalyzes the condensation of homocysteine and serine to form cystathionine, with the release of water. In humans, deficiency in CBS activity is the most common cause of hyperhomocysteinaemia and homocystinuria. More than 160 pathogenic mutations in the human CBS gene have been described to date. Here, the purification and preliminary crystallographic analysis of the catalytic core of CBS from Saccharomyces cerevisiae (ScCBS) is described which, in contrast to other eukaryotic CBSs, lacks the N-terminal haem-binding domain and is considered to be a useful model for investigation of the pyridoxal-5'-phosphate-mediated reactions of human CBS (hCBS). The purified protein yielded two different crystal forms belonging to space groups P41212 and P212121, with unit-cell parameters a = b = 72.390, c = 386.794 Å and a = 58.156, b = 89.988, c = 121.687 Å, respectively. Diffraction data were collected to 2.7 and 3.1 Å resolution, respectively, using synchrotron radiation. Preliminary analysis of the X-ray data suggests the presence of ScCBS homodimers in both types of crystals.

Entities:  

Keywords:  CBS domain; S-adenosylmethionine; cystathionine β-synthase; cysteine biosynthesis; homocysteine; pyridoxal-5′-phosphate; transsulfuration pathway

Mesh:

Substances:

Year:  2014        PMID: 24598918      PMCID: PMC3944693          DOI: 10.1107/S2053230X14001502

Source DB:  PubMed          Journal:  Acta Crystallogr F Struct Biol Commun        ISSN: 2053-230X            Impact factor:   1.056


  35 in total

1.  Transsulfuration in Saccharomyces cerevisiae is not dependent on heme: purification and characterization of recombinant yeast cystathionine beta-synthase.

Authors:  K N Maclean; M Janosík; J Oliveriusová; V Kery; J P Kraus
Journal:  J Inorg Biochem       Date:  2000-08-31       Impact factor: 4.155

2.  Impaired heme binding and aggregation of mutant cystathionine beta-synthase subunits in homocystinuria.

Authors:  M Janosík; J Oliveriusová; B Janosíková; J Sokolová; E Kraus; J P Kraus; V Kozich
Journal:  Am J Hum Genet       Date:  2001-05-15       Impact factor: 11.025

3.  Plasma homocysteine as a risk factor for dementia and Alzheimer's disease.

Authors:  Sudha Seshadri; Alexa Beiser; Jacob Selhub; Paul F Jacques; Irwin H Rosenberg; Ralph B D'Agostino; Peter W F Wilson; Philip A Wolf
Journal:  N Engl J Med       Date:  2002-02-14       Impact factor: 91.245

4.  Characterization of transsulfuration and cysteine biosynthetic pathways in the protozoan hemoflagellate, Trypanosoma cruzi. Isolation and molecular characterization of cystathionine beta-synthase and serine acetyltransferase from Trypanosoma.

Authors:  T Nozaki; Y Shigeta; Y Saito-Nakano; M Imada; W D Kruger
Journal:  J Biol Chem       Date:  2000-12-05       Impact factor: 5.157

5.  Regulation of human cystathionine beta-synthase by S-adenosyl-L-methionine: evidence for two catalytically active conformations involving an autoinhibitory domain in the C-terminal region.

Authors:  M Janosík; V Kery; M Gaustadnes; K N Maclean; J P Kraus
Journal:  Biochemistry       Date:  2001-09-04       Impact factor: 3.162

6.  Structure of human cystathionine beta-synthase: a unique pyridoxal 5'-phosphate-dependent heme protein.

Authors:  M Meier; M Janosik; V Kery; J P Kraus; P Burkhard
Journal:  EMBO J       Date:  2001-08-01       Impact factor: 11.598

7.  The reaction of yeast cystathionine beta-synthase is rate-limited by the conversion of aminoacrylate to cystathionine.

Authors:  K H Jhee; D Niks; P McPhie; M F Dunn; E W Miles
Journal:  Biochemistry       Date:  2001-09-11       Impact factor: 3.162

8.  Stopped-flow kinetic analysis of the reaction catalyzed by the full-length yeast cystathionine beta-synthase.

Authors:  Shinichi Taoka; Ruma Banerjee
Journal:  J Biol Chem       Date:  2002-04-10       Impact factor: 5.157

Review 9.  From cofactor to enzymes. The molecular evolution of pyridoxal-5'-phosphate-dependent enzymes.

Authors:  P Christen; P K Mehta
Journal:  Chem Rec       Date:  2001       Impact factor: 6.771

10.  Yeast cystathionine beta-synthase is a pyridoxal phosphate enzyme but, unlike the human enzyme, is not a heme protein.

Authors:  K H Jhee; P McPhie; E W Miles
Journal:  J Biol Chem       Date:  2000-04-21       Impact factor: 5.157

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