Calcium dependent formation of tubular assemblies by recombinant S-layer proteins in vivo and in vitro.
Surface layer proteins have the appealing property to self-assemble in nanosized arrays in solution and on solid substrates. In this work, we characterize the formation of assembly structures of the recombinant surface layer protein SbsC of Geobacillus stearothermophilus ATTC 12980, which was tagged with enhanced green fluorescent protein and expressed in the yeast Saccharomyces cerevisiae. The tubular structures formed by the protein in vivo are retained upon bursting the cells by osmotic shock; however, their average length is decreased. During dialysis, monomers obtained by treatment with chaotropic chemicals recrystallize again to form tube-like structures. This process is strictly dependent on calcium (Ca(2+)) ions, with an optimal concentration of 10 mM. Further increase of the Ca(2+) concentration results in multiple non-productive nucleation points. We further show that the lengths of the S-layer assemblies increase with time and can be controlled by pH. After 48 h, the average length at pH 9.0 is 4.13 µm compared to 2.69 µm at pH 5.5. Successful chemical deposition of platinum indicates the potential of recrystallized mSbsC-eGFP structures for nanobiotechnological applications.
Institut für Genetik, Technische Universität Dresden, Dresden, Germany. nuriye_korkmaz@yahoo.com
Nuriye
N
Korkmaz
Kai
K
Ostermann
Gerhard
G
Rödel
D002118:Calcium / Q000737:chemistry* / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
D008562:Membrane Glycoproteins; D046912:Multiprotein Complexes; D011994:Recombinant Proteins; C055764:S-layer proteins; D049452:Green Fluorescent Proteins; D002118:Calcium