The reconstitution of active holoenzyme containing calcium from inactive calcium-free methanol dehydrogenase, isolated from a moxA mutant of Methylobacterium extorquens, has a pH optimum of about pH 10, with a well defined pK for the process at pH 9.3. Two Ca2+ ions were irreversibly incorporated per α2β2 tetramer. Calcium could be replaced in the incorporation process by strontium or barium, the affinities for these ions being similar to that for Ca2+. Arrhenius plots for measurement of the activation energy of reconstitution were biphasic; the lower activation energy was typical of most biological processes, while the higher activation energy was at least three times greater, implying the involvement of a large conformational change during incorporation of the cations. The activation energy for incorporation of Ba2+ was considerably higher than that for incorporation of Ca2+. The novel disulphide bridge that is at the active site of the enzyme was not involved in the incorporation process. Studies of the time courses for incorporation of 45Ca2+, production of active enzyme and changes in absorption spectra failed to show any intermediates in the incorporation process.
Skip Nav Destination
Article navigation
November 1996
-
Cover Image
Cover Image
- PDF Icon PDF LinkTable of Contents
Research Article|
November 01 1996
Reconstitution of the quinoprotein methanol dehydrogenase from inactive Ca2+-free enzyme with Ca2+, Sr2+ or Ba2+
Matthew G GOODWIN;
Matthew G GOODWIN
1Department of Biochemistry, School of Biological Sciences, University of Southampton, Southampton SO16 7PX, U.K.
Search for other works by this author on:
Alain AVEZOUX;
Alain AVEZOUX
1Department of Biochemistry, School of Biological Sciences, University of Southampton, Southampton SO16 7PX, U.K.
Search for other works by this author on:
Simon L DALES;
Simon L DALES
1Department of Biochemistry, School of Biological Sciences, University of Southampton, Southampton SO16 7PX, U.K.
Search for other works by this author on:
Christopher ANTHONY
Christopher ANTHONY
*
1Department of Biochemistry, School of Biological Sciences, University of Southampton, Southampton SO16 7PX, U.K.
*To whom correspondence should be addressed.
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
May 15 1996
Revision Received:
July 10 1996
Accepted:
July 12 1996
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1996
1996
Biochem J (1996) 319 (3): 839–842.
Article history
Received:
May 15 1996
Revision Received:
July 10 1996
Accepted:
July 12 1996
Citation
Matthew G GOODWIN, Alain AVEZOUX, Simon L DALES, Christopher ANTHONY; Reconstitution of the quinoprotein methanol dehydrogenase from inactive Ca2+-free enzyme with Ca2+, Sr2+ or Ba2+. Biochem J 1 November 1996; 319 (3): 839–842. doi: https://doi.org/10.1042/bj3190839
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.