Electron microscopy and computational studies of Ebh, a giant cell-wall-associated protein from Staphylococcus aureus.
Ebh, a giant protein found in staphylococci, contains several domains, including a large central region with 52 imperfect repeats of a domain composed of 126 amino acids. We used electron microscopy to observe the rod-like structure of a partial Ebh protein containing 10 repeating units. This is the first report of the direct observation of an Ebh structure containing a large number of repeating units, although structures containing one, two, or four repeating units have been reported. The observed structure of the partial Ebh protein was distorted and had a length of ca. 520A and a width of ca. 21A. The observed structures were consistent with those deduced from crystal structure analysis, suggesting that the Ebh domains are connected to form a rod-like structure. The crystal structure data revealed distorted, string-like features in the simulated structure of the whole-length Ebh protein. Superposition of fragments of the simulated whole-length structure of the Ebh protein onto each electron micrograph showed a high level of correlation between the observed and calculated structures. These results suggest that Ebh is composed of highly flexible filate molecules. The highly repetitive structure and the associated unique structural flexibility of Ebh support the proposed function of this protein, i.e. binding to sugars in the cell wall. This binding might result in intra-cell-wall cross-linking that contributes to the rigidity of bacterial cells.
Sakamoto
Sou
S
Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, 301 FBS-Building, 5-1-5 Kashiwanoha, Kashiwa 277-8562, Japan.
Tanaka
Yoshikazu
Y
Tanaka
Isao
I
Takei
Toshiaki
T
Yu
Jian
J
Kuroda
Makoto
M
Yao
Min
M
Ohta
Toshiko
T
Tsumoto
Kouhei
K
D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics; D019295:Computational Biology; D008854:Microscopy, Electron; D008958:Models, Molecular; D011487:Protein Conformation; D013211:Staphylococcus aureus / Q000378:metabolism*
D001426:Bacterial Proteins; D002352:Carrier Proteins; C469917:Ebh protein, Staphylococcus aureus