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Content archived on 2024-04-15

PROTEIN DESIGN AND NEW CATALYTIC MECHANISMS WITH PYRROLOQUINOLINE QUINONE PROSTHETIC GROUP

Objective


Methylamine dehydrogenase from Thiobacillus versutus was obtained as a homogeneous preparation. From the molecular weight and studies on the subunits, it appears that it has an alpha 2 beta 2 structure. Homogeneous preparations of the alpha and beta subunits were obtained and by using specific proteases, peptides prepared by high pressure liquid chromatography. A number of peptides were sequenced, providing information to construct probes to clone the genes, to elucidate the amino acid sequence around the attachment sites of the cofactor to the protein chain, and to support the work on the 3-dimensional structure elucidation. Results prove that the cofactor in the enzyme from Th versutus is also tryptophan tryptophyl quinone (TTQ). Attempts to crystallyze the enzyme were successful which led to the first 3-dimensional structure elucidation of a quinoprotein enzyme. The use of crystals treated with inhibitors has shown that the reactive carbonyl group is situated at the C6 carbon atom.

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RIJKSUNIVERSITEIT GRONINGEN
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P.B.72
9700 AB GRONINGEN
Netherlands

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