The pyruvate formate lyase family: sequences, structures and activation.
We cloned and expressed in Escherichia coli the Archaeglobus fulgidus gene that encodes pyruvate formate lyase 2 (PFL2). PFL2, despite its homology to the other glycyl radical enzymes, differs from them by exhibiting a completely different oligomerization. The most abundant form of PFL2 when expressed in E.coli is a trimer. The closest homologue of PFL2 with a known structure is E. coli PFL, which is a dimer. Sequence comparisons allowed us to reclassify PFL-like enzymes and the consensus sequences allowed us to propose an activation route for PFL-like glycyl radical enzymes. Surprisingly, most of the conserved residues in PFL-like enzymes appear to be involved in preserving the structure, rather than forming the active site.
Lehtiö
L
L
Graduate School in Informational and Structural Biology, Institute of Biotechnology, University of Helsinki, PO Box 65, FIN-00014 Helsinki, Finland.
Goldman
A
A
D000123:Acetyltransferases / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D000595:Amino Acid Sequence; D019611:Archaeoglobus fulgidus / Q000201:enzymology* / Q000235:genetics; D020134:Catalytic Domain / Q000235:genetics; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D016384:Consensus Sequence / Q000235:genetics; D017124:Conserved Sequence / Q000235:genetics; D003545:Cysteine / Q000235:genetics; D019641:DNA, Archaeal / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics; D008027:Light; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D012542:Scattering, Radiation; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D019641:DNA, Archaeal; D021122:Protein Subunits; D011994:Recombinant Proteins; D000123:Acetyltransferases; C022560:formate C-acetyltransferase; D003545:Cysteine