Advanced

23S rRNA assisted folding of cytoplasmic malate dehydrogenase is distinctly different from its self-folding

Sanyal, Suparna LU ; Pal, S; Chaudhuri, S and DasGupta, C (2002) In Nucleic Acids Research 30(11). p.2390-2397
Abstract
The role of the 50S particle of Escherichia coli ribosome and its 23S rRNA in the refolding and subunit association of dimeric porcine heart cytoplasmic malate dehydrogenase (s-MDH) has been investigated. The self-reconstitution of s-MDH is governed by two parallel pathways representing the folding of the inactive monomeric and the dimeric intermediates. However, in the presence of these folding modulators, only one first order kinetics was observed. To understand whether this involved the folding of the monomers or the dimers, subunit association of s-MDH was studied using fluorescein-5-isothiocyanate-rhodamine-isothiocyanate (FITC-RITC) fluorescence energy transfer and chemical crosslinking with gluteraldehyde. The observation suggests... (More)
The role of the 50S particle of Escherichia coli ribosome and its 23S rRNA in the refolding and subunit association of dimeric porcine heart cytoplasmic malate dehydrogenase (s-MDH) has been investigated. The self-reconstitution of s-MDH is governed by two parallel pathways representing the folding of the inactive monomeric and the dimeric intermediates. However, in the presence of these folding modulators, only one first order kinetics was observed. To understand whether this involved the folding of the monomers or the dimers, subunit association of s-MDH was studied using fluorescein-5-isothiocyanate-rhodamine-isothiocyanate (FITC-RITC) fluorescence energy transfer and chemical crosslinking with gluteraldehyde. The observation suggests that during refolding the interaction of the unstructured monomers of s-MDH with these ribosomal folding modulators leads to very fast formation of structured monomers that immediately dimerise. These inactive dimers then fold to the native ones, which is the rate limiting step in 23S or 50S assisted refolding of s-MDH. Furthermore, the sequential action of the two fragments of domain V of 23S rRNA has been investigated in order to elucidate the mechanism. The central loop of domain V of 23S rRNA (RNA1) traps the monomeric intermediates, and when they are released by the upper stem-loop region of the domain V of 23S rRNA (RNA2) they are already structured enough to form dimeric intermediates which are directed towards the proper folding pathway. (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
Nucleic Acids Research
volume
30
issue
11
pages
2390 - 2397
publisher
Oxford University Press
external identifiers
  • wos:000176256500015
  • pmid:12034826
ISSN
1362-4962
DOI
10.1093/nar/30.11.2390
language
English
LU publication?
yes
id
ebde673d-53d5-4897-bf48-998df5d83745 (old id 334734)
date added to LUP
2007-11-12 15:31:48
date last changed
2016-04-15 20:04:36
@article{ebde673d-53d5-4897-bf48-998df5d83745,
  abstract     = {The role of the 50S particle of Escherichia coli ribosome and its 23S rRNA in the refolding and subunit association of dimeric porcine heart cytoplasmic malate dehydrogenase (s-MDH) has been investigated. The self-reconstitution of s-MDH is governed by two parallel pathways representing the folding of the inactive monomeric and the dimeric intermediates. However, in the presence of these folding modulators, only one first order kinetics was observed. To understand whether this involved the folding of the monomers or the dimers, subunit association of s-MDH was studied using fluorescein-5-isothiocyanate-rhodamine-isothiocyanate (FITC-RITC) fluorescence energy transfer and chemical crosslinking with gluteraldehyde. The observation suggests that during refolding the interaction of the unstructured monomers of s-MDH with these ribosomal folding modulators leads to very fast formation of structured monomers that immediately dimerise. These inactive dimers then fold to the native ones, which is the rate limiting step in 23S or 50S assisted refolding of s-MDH. Furthermore, the sequential action of the two fragments of domain V of 23S rRNA has been investigated in order to elucidate the mechanism. The central loop of domain V of 23S rRNA (RNA1) traps the monomeric intermediates, and when they are released by the upper stem-loop region of the domain V of 23S rRNA (RNA2) they are already structured enough to form dimeric intermediates which are directed towards the proper folding pathway.},
  author       = {Sanyal, Suparna and Pal, S and Chaudhuri, S and DasGupta, C},
  issn         = {1362-4962},
  language     = {eng},
  number       = {11},
  pages        = {2390--2397},
  publisher    = {Oxford University Press},
  series       = {Nucleic Acids Research},
  title        = {23S rRNA assisted folding of cytoplasmic malate dehydrogenase is distinctly different from its self-folding},
  url          = {http://dx.doi.org/10.1093/nar/30.11.2390},
  volume       = {30},
  year         = {2002},
}