Intermediate filament-like network formed in vitro by a bacterial coiled coil protein.
The TlpA protein encoded by the virulence plasmid of Salmonella enterica is an alpha-helical 371-amino acid protein possessing characteristics similar to eukaryotic coiled coil proteins (Koski, P., Saarilahti, H., Sukupolvi, S., Taira, S., Rikkonen, P., Osterlund, K., Hurme, R., and Rhen, M. (1992) J. Biol. Chem. 267, 12258-12265). In this paper we have investigated inter- and intramolecular associations and the morphology of structures formed by TlpA. Dynamics and temperature stability of TlpA dimers were studied by examining the feasibility and conditions in which TlpA would form an artificial heterodimer with its truncated derivative. Formation of heterodimers, bridged by Cu(2+)-catalyzed air oxidation of adjacent Cys residues, showed that TlpA dimers are dynamic chain exchanging structures at 37 degrees C, whereas they were nonexchanging at room temperature or on ice. Chemical cross-linking suggested higher order interaction between TlpA dimers. Electron microscopy studies revealed two levels of TlpA organization in vitro: thin filaments and rods, 2-5 nm in diameter, and a higher ordered filament network consisting of tonofilament-like formations with a diameter of 8-15 nm. Electron microscopy of thin-sectioned Escherichia coli over-producing TlpA showed an extraordinary intracellular assembly of proteinacious lamellae with a striated appearance and a 38-nm periodicity. This study describes for the first time a bacterial protein capable of organizing itself into an ordered and suspectedly dynamic intermediate filament-like architecture.
Hurme
R
R
Department of Biochemistry, University of Helsinki, Finland.
Namork
E
E
Nurmiaho-Lassila
E L
EL
Rhen
M
M
D000388:Air; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D001483:Base Sequence; D003432:Cross-Linking Reagents; D004926:Escherichia coli / Q000235:genetics / Q000648:ultrastructure; D007381:Intermediate Filament Proteins / Q000737:chemistry*; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010084:Oxidation-Reduction; D011108:Polymers; D012475:Salmonella
D001426:Bacterial Proteins; D003432:Cross-Linking Reagents; D007381:Intermediate Filament Proteins; D009838:Oligodeoxyribonucleotides; D011108:Polymers; C075138:TlpA protein, Salmonella