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End Group Analysis of the Cytosolic and Mitochondrial Fumarases from Rat Liver

Some molecular properties of the cytosolic and mitochondrial fumarases were compared. The carboxyl(C)-terminal amino acid of both the cytosolic and mitochondrial fumarases of rat liver cell was identified as leucine by using carboxypep-tidase (CPase) A. As the amino(N)-terminal amino acid of both th...

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Bibliographic Details
Published in:Journal of biochemistry (Tokyo) 1983-01, Vol.94 (3), p.707-713
Main Authors: KOBAYASHI, Keiko, TUBOI, Syozo
Format: Article
Language:English
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Summary:Some molecular properties of the cytosolic and mitochondrial fumarases were compared. The carboxyl(C)-terminal amino acid of both the cytosolic and mitochondrial fumarases of rat liver cell was identified as leucine by using carboxypep-tidase (CPase) A. As the amino(N)-terminal amino acid of both the cytosolic and mitochondrial fumarases could not be identified by the dansyl chloride method or by the cyanate method, the N-termini of these two fumarases seems to be masked. Both fumarases, after S-carboxymethylation, were completely digested with pronase E and CPase A and B, and the amino acids with blocked amino group were analyzed by high voltage paper electrophoresis and amino acid analysis after acid hydrolysis of these amino acid derivatives. The N-termini of the mitochondrial and cytosolic fumarases were identified as pyroglutamic acid and N-acetylalanine, respectively. To compare the primary structures of the two fumarases in detail, each fuma-rase was digested with an arginine-specific protease or cleaved with cyanogen bromide. The elecrrophoretic profiles of the digests of these fumarases were indistinguishable from each other.
ISSN:0021-924X
DOI:10.1093/oxfordjournals.jbchem.a134410