A yeast system for expression of human cystathionine beta-synthase: structural and functional conservation of the human and yeast genes.
Human cystathionine beta-synthase (CBS; EC 4.2.1.22) deficiency results in a recessive genetic disorder whose clinical and biochemical manifestations vary greatly among affected individuals. In an effort to identify and analyze mutations in the human CBS gene, we have developed a yeast expression system for human CBS. We have cloned and sequenced a human cDNA that codes for CBS and have expressed the human CBS protein in yeast cells lacking endogenous CBS. The human enzyme produced in yeast is functional both in vitro and in vivo. We have also cloned and sequenced the yeast gene, CYS4, that codes for CBS. The predicted human and yeast CBS proteins are 38% identical and 72% similar to each other, as well as sharing significant similarity with bacterial cysteine synthase. These results demonstrate the evolutionary conservation of CBS and establish the utility of a yeast expression system for studying human CBS.
Department of Psychiatry, University of California at San Francisco 94143.
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D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D003541:Cystathionine beta-Synthase / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015723:Gene Library; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D015186:Hominidae / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D011994:Recombinant Proteins; D003541:Cystathionine beta-Synthase