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  Novel molecular architecture of the multimeric archaeal PEP-synthase homologue (MAPS) from Staphylothermus marinus

Cicicopol, C., Peters, J., Lupas, A., Cejka, Z., Müller, S. A., Golbik, R., et al. (1999). Novel molecular architecture of the multimeric archaeal PEP-synthase homologue (MAPS) from Staphylothermus marinus. Journal of Molecular Biology, 290(1), 347-361.

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Genre: Journal Article
Alternative Title : J. Mol. Biol

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Cicicopol, C., Author
Peters, J.1, Author              
Lupas, A.1, Author              
Cejka, Z., Author
Müller, S. A., Author
Golbik, R., Author
Pfeifer, G.1, Author              
Lilie, H., Author
Engel, A., Author
Baumeister, W.1, Author              
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1External Organizations, ou_persistent22              

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Free keywords: Pep-synthase; Archaea; Homomultimer; Coiled-coil; Oligomerization domain.; Homomultimeric phosphoenolpyruvate synthase; Dehydrogenase multienzyme complex; Transmission electron-microscopy; Oligomeric matrix protein; Alpha-helical bundle; Escherichia-coli; Pyrococcus-furiosus; Crystal-structure; Coiled-coils; Gene.; Molecular Biology & Genetics in Current Contents(R)/Life Sciences.
 Abstract: The phosphoenolpyruvate (PEP)-synthases belong to the family of structurally and functionally related PEP-utilizing enzymes. The only archaeal member of this family characterized thus far is the Multimeric Archaeal PEP-Synthase homologue from Staphylothermus marinus (MAPS). This protein complex differs from the bacterial and eukaryotic representatives characterized to date in its homomultimeric, as opposed to dimeric or tetrameric, structure. We have probed the molecular architecture of MAPS using limited proteolytic digestion in conjunction with electron microscopic, biochemical, and biophysical techniques. The 2.2 MDa particle was found to be organized! in a concentric fashion. The 93.7 kDa monomers possess a pronounced tripartite domain structure and are arranged such that the N-terminal domains form an outer shell, the intermediate domains form an inner shell, and the C-terminal domains form a core structure responsible for the assembly into a multimeric complex. The core domain was shown to be capable of assembling into the native multimer by recombinant expression in Escherichia coli. Deletion mutants as well as a synthetic peptide were investigated for their state of oligomerization using native polyacrylamide gel electrophoresis, molecular sieve chromatography, analytical ultracentrifugation, circular dichroism (CD) spectroscopy, and chemical cross-linking. Our data confirmed the existence of a short C-terminal, tx-helical oligomerization motif that had been suggested by multiple sequence alignments and secondary structure predictions. We propose that this motif bundles the monomers into six groups of four. An additional formation of 12 dimers between globular domains from different bundles leads to the multimeric assembly. According to our model, each of the six bundles of globular domains is positioned at the corners of an imaginary octahedron, and the helical C-terminal segments are oriented towards the centre of the particle. The edges of the octahedron represent the dimeric contacts. Phylogenetic analysis suggests that the ancient predecessor of this family of enzymes contained the C-terminal oligomerization motif as a feature that was preserved in some hyperthermophiles. (C) 1999 Academic Press. [References: 33]

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 Dates: 1999
 Publication Status: Published in print
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 Identifiers: eDoc: 318644
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Title: Journal of Molecular Biology
  Alternative Title : J. Mol. Biol
Source Genre: Journal
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Pages: - Volume / Issue: 290 (1) Sequence Number: - Start / End Page: 347 - 361 Identifier: -