This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Kiszkiss, D. F. Articles by Downey, R. J. Search for Related Content PubMed PubMed Citation Articles by Kiszkiss, D. F. Articles by Downey, R. J.

 Previous Article  |  Next Article 

J Bacteriol. 1972 February; 109(2): 803-810
Copyright © 1972 American Society for Microbiology. All Rights Reserved.

Localization and Solubilization of the Respiratory Nitrate Reductase of Bacillus stearothermophilus¹

^a Department of Microbiology, University of Notre Dame, Notre Dame, Indiana 46556

ABSTRACT

Membranes were isolated from Bacillus stearothermophilus 2184D by lysozyme digestion of the cell wall and subsequent differential centrifugation. Observations with the electron microscope indicate that such membranes are relatively intact and have a typical membrane appearance. Nitrate will preferentially oxidize the cytochrome b of such membranes. Approximately 80% of the total respiratory nitrate reductase activity of whole cells can be localized in the washed membrane fraction and the process of membrane isolation results in a sixfold purification of this enzyme. Of several detergents tested, sodium dodecyl sulfate, Triton 114, and Triton X-100 are most effective in converting reduced methyl viologen-nitrate reductase to a form which will not pellet at 130,000 x g. Density gradient analysis reveals that such detergent-mediated solubilization converts virtually all membrane protein to a form of lighter density.

¹ Part of the dissertation of D. F. Kiszkiss, presented to the Graduate School of Notre Dame University in partial fulfillment of requirements for the Ph.D. degree.