Despite great interest in their remarkable properties, and the considerable range of expertise brought to bear by several groups over many years, only the most general features of the structure of gas vesicles are known. However, recent developments now poise solid state NMR to provide numerous unique probes of the atomic details of the structure of the 7kDa monomer, gvpA, and its assembly into rigid, amyloid-like shells. Preliminary 13C and 15N spectra of uniformly labeled vesicles indicate that a combination of several isotope labeling strategies and an array of multidimensional correlation experiments should allow the full assignment of resonances and the determination of both intra-monomer and inter-monomer structural constraints. Ultimately, this information will allow the complete structure of the gas vesicle shell to be determined. Work will be carried out simultaneously on gas vesicles from the archae Halobacterium salinarum and the cyanobacterium Anabaena flos-aquaea. The parallel efforts will be synergistic because of the strong core homology between the gvpA's of the two species and the different isotope labeling strategies available in the two organisms. The parallel efforts will also be complimentary because of differences in overall vesicle morphology associated with differences in the N- and C-terminal domains of the two gvpA sequences. The role of electrostatics will be explored by identifying the cation displacements and protonation changes that induce vesicle collapse with decreasing pH. The response to salt will also be interesting since H. salinarum is a halophile and A. flos-aquaea is adapted to sweet water. Discovered structural details will shed light on the molecular basis of the morphology and rigidity of the vesicles, the prevention of water condensation within these gas permeable organelles, and the controlled assembly and disassembly of these amyloid-like aggregates.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
5R01EB002175-03
Application #
6856474
Study Section
Biophysical Chemistry Study Section (BBCB)
Program Officer
Mclaughlin, Alan Charles
Project Start
2003-05-01
Project End
2008-02-29
Budget Start
2005-03-01
Budget End
2006-02-28
Support Year
3
Fiscal Year
2005
Total Cost
$303,234
Indirect Cost
Name
Brandeis University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
616845814
City
Waltham
State
MA
Country
United States
Zip Code
02454
Herzfeld, Judith; Rand, Danielle; Matsuki, Yoh et al. (2011) Molecular structure of humin and melanoidin via solid state NMR. J Phys Chem B 115:5741-5
Sivertsen, Astrid C; Bayro, Marvin J; Belenky, Marina et al. (2010) Solid-state NMR characterization of gas vesicle structure. Biophys J 99:1932-9
Sivertsen, Astrid C; Bayro, Marvin J; Belenky, Marina et al. (2009) Solid-state NMR evidence for inequivalent GvpA subunits in gas vesicles. J Mol Biol 387:1032-9
Mamajanov, Irena; Herzfeld, Judith (2009) HCN polymers characterized by SSNMR: solid state reaction of crystalline tetramer (diaminomaleonitrile). J Chem Phys 130:134504
Mamajanov, Irena; Herzfeld, Judith (2009) HCN polymers characterized by solid state NMR: chains and sheets formed in the neat liquid. J Chem Phys 130:134503