Macromolecular organization of the Yersinia pestis capsular F1 antigen: insights from time-of-flight mass spectrometry.
Mass spectrometry has been used to examine the subunit interactions in the capsular F1 antigen from Yersinia pestis, the causative agent of the plague. Introducing the sample using nanoflow electrospray from solution conditions in which the protein remains in its native state and applying collisional cooling to minimize the internal energy of the ions, multiple subunit interactions have been maintained. This methodology revealed assemblies of the F1 antigen that correspond in mass to both 7-mers and 14-mers, consistent with interaction of two seven-membered units. The difference between the calculated masses and those measured experimentally for these higher-order oligomers was found to increase proportionately with the size of the complex. This is consistent with a solvent-filled central cavity maintained on association of the 7-mer to the 14-mer. The charge states of the ions show that an average of one and four surface accessible basic side-chains are involved in maintaining the interactions between the 7-mer units and neighboring subunits, respectively. Taken together, these findings provide new information about the stoichiometry and packing of the subunits involved in the assembly of the capsular antigen structure. More generally, the data show that the symmetry and packing of macromolecular complexes can be determined solely from mass spectrometry, without any prior knowledge of higher order structure
Tito
M A
MA
Oxford Centre for Molecular Sciences, New Chemistry Laboratory, Oxford, OX1 3QT, United Kingdom.
Miller
J
J
Griffin
K F
KF
Williamson
E D
ED
Titball
R W
RW
Robinson
C V
CV
D016667:Bacterial Capsules / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods; D008956:Models, Chemical; D008958:Models, Molecular; D012996:Solutions; D015010:Yersinia pestis / Q000737:chemistry* / Q000276:immunology
D001426:Bacterial Proteins; D012996:Solutions; C086725:caf1 protein, Yersinia pestis