Advanced

In Vitro Assembly of Catalase.

Baureder, Michael LU ; Barane, Elisabeth LU and Hederstedt, Lars LU (2014) In Journal of Biological Chemistry 289(41). p.28411-28420
Abstract
Most aerobic organisms contain catalase which functions to decompose hydrogen peroxide. Typical catalases are structurally complex homo-tetrameric enzymes with one heme prosthetic group buried in each subunit. It is not known how catalase in the cell is assembled from its constituents. The bacterium Enterococcus faecalis cannot synthesize heme but can acquire it from the environment to form a cytoplasmic catalase. We have in E. faecalis monitored production of the enzyme polypeptide (KatA) depending on the availability of heme and used our findings to devise a procedure for the purification of preparative amounts of in vivo-synthesized apocatalase. We show that fully active catalase can be obtained in vitro by incubating isolated... (More)
Most aerobic organisms contain catalase which functions to decompose hydrogen peroxide. Typical catalases are structurally complex homo-tetrameric enzymes with one heme prosthetic group buried in each subunit. It is not known how catalase in the cell is assembled from its constituents. The bacterium Enterococcus faecalis cannot synthesize heme but can acquire it from the environment to form a cytoplasmic catalase. We have in E. faecalis monitored production of the enzyme polypeptide (KatA) depending on the availability of heme and used our findings to devise a procedure for the purification of preparative amounts of in vivo-synthesized apocatalase. We show that fully active catalase can be obtained in vitro by incubating isolated apoprotein with hemin. We have characterized features of the assembly process and describe a temperature-trapped hemylated intermediate of the enzyme maturation process. Hemylation of apocatalase does not require auxiliary cell components but rapid assembly of active enzyme seemingly is assisted in the cell. Our findings provide insight about catalase assembly and offer new experimental possibilities for detailed studies of this process. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biological Chemistry
volume
289
issue
41
pages
28411 - 28420
publisher
ASBMB
external identifiers
  • pmid:25148685
  • wos:000343765400034
  • scopus:84907821989
ISSN
1083-351X
DOI
10.1074/jbc.M114.596148
language
English
LU publication?
yes
id
498ff559-0c9e-41ae-b698-da05b610e657 (old id 4614160)
date added to LUP
2014-09-10 15:57:45
date last changed
2017-01-29 03:06:31
@article{498ff559-0c9e-41ae-b698-da05b610e657,
  abstract     = {Most aerobic organisms contain catalase which functions to decompose hydrogen peroxide. Typical catalases are structurally complex homo-tetrameric enzymes with one heme prosthetic group buried in each subunit. It is not known how catalase in the cell is assembled from its constituents. The bacterium Enterococcus faecalis cannot synthesize heme but can acquire it from the environment to form a cytoplasmic catalase. We have in E. faecalis monitored production of the enzyme polypeptide (KatA) depending on the availability of heme and used our findings to devise a procedure for the purification of preparative amounts of in vivo-synthesized apocatalase. We show that fully active catalase can be obtained in vitro by incubating isolated apoprotein with hemin. We have characterized features of the assembly process and describe a temperature-trapped hemylated intermediate of the enzyme maturation process. Hemylation of apocatalase does not require auxiliary cell components but rapid assembly of active enzyme seemingly is assisted in the cell. Our findings provide insight about catalase assembly and offer new experimental possibilities for detailed studies of this process.},
  author       = {Baureder, Michael and Barane, Elisabeth and Hederstedt, Lars},
  issn         = {1083-351X},
  language     = {eng},
  number       = {41},
  pages        = {28411--28420},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {In Vitro Assembly of Catalase.},
  url          = {http://dx.doi.org/10.1074/jbc.M114.596148},
  volume       = {289},
  year         = {2014},
}