Components of the regular surface array of Aquaspirillum serpens MW5 and their assembly in vitro

Abstract

The two-layered regular surface array of Aquaspirillum serpens MW5 was removed from cell envelopes and dissociated into subunits by treatment with 6 M urea. The surface components reassembled onto an outer membrane surface and self-assembled into planar sheets in vitro in the presence of Ca2+ or Sr2+. The two layers were removed sequentially from cell envelopes by a two-step extraction procedure involving initial treatment with a high-pH buffer to remove the outermost surface layer and subsequent treatment with 6 M urea to remove the innermost layer. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the outer and inner layers of the array were composed of two proteins with molecular weights of 125,000 and 150,000, respectively. The two layers assembled sequentially; the 150,000-molecular-weight protein formed an array on an outer membrane surface, and the 125,000-molecular-weight protein required that array as a template for its in vitro assembly.