ISSN:
1432-0983
Keywords:
CBS1 protein
;
Protein purification
;
Mitochondrial presequence
;
Protein sequencing
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Summary Yeast transformats harboring the CBS1 gene under the control of the strong ADC1 promoter on a high copy number plasmid express the mitochondrial CBS1 protein at artificially high levels. Over-expressed protein is imported into mitochondria and correctly processed to vield the mature mitochondrial 23.5 kDa form, but differs in its solubility properties from CBS1 in wild-type mitochondria. It forms insoluble protein aggregates, which are refractory to solubilization with 1% Taurodeoxycholate. We exploited this observation to separate CBS1 from the bulk of mitochondrial proteins and to isolate CBS1 after SDS gel electrophoresis. Determination of the amino-terminal amino acids of the purified protein reveals that the mature CBS1 protein starts with Ile30, at the characteristic distance of +2 amino acids from an arginine residue (Arg28). The cleavage site shows a remarkable homology to that of subunit 9 of the F0F1 ATPase from Neurospora crassa.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00312770
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