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. 1988 Sep 1;254(2):427–435. doi: 10.1042/bj2540427

Purification, crystallization and properties of porphobilinogen deaminase from a recombinant strain of Escherichia coli K12.

P M Jordan 1, S D Thomas 1, M J Warren 1
PMCID: PMC1135095  PMID: 3052434

Abstract

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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