Porphobilinogen deaminase has been purified and crystallized from an overproducing recombinant strain of Escherichia coli harbouring a hemC-containing plasmid which has permitted the purification of milligram quantities of the enzyme. Determination of the Mr of the enzyme by SDS/polyacrylamide-gel electrophoresis (35,000) and gel filtration (32,000) agrees with the gene-derived Mr of 33,857. The enzyme has a Km of 19 +/- 7 microM, an isoelectric point of 4.5 and an N-terminal sequence NH2-MLDNVLRIAT. The substrate, porphobilinogen, binds to the active-site dipyrromethane cofactor to form three intermediate complexes: ES, ES2 and ES3. The gene-derived primary structure of the E. coli deaminase is compared with that derived from the cDNA of the human enzyme.
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September 1988
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Research Article|
September 01 1988
Purification, crystallization and properties of porphobilinogen deaminase from a recombinant strain of Escherichia coli K12
P M Jordan;
P M Jordan
Department of Biochemistry, University of Southampton, Southampton S09 3TU, U.K.
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S D Thomas;
S D Thomas
Department of Biochemistry, University of Southampton, Southampton S09 3TU, U.K.
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M J Warren
M J Warren
Department of Biochemistry, University of Southampton, Southampton S09 3TU, U.K.
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Publisher: Portland Press Ltd
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 1988 London: The Biochemical Society
1988
Biochem J (1988) 254 (2): 427–435.
Citation
P M Jordan, S D Thomas, M J Warren; Purification, crystallization and properties of porphobilinogen deaminase from a recombinant strain of Escherichia coli K12. Biochem J 1 September 1988; 254 (2): 427–435. doi: https://doi.org/10.1042/bj2540427
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