Advanced

Inhibition kinetics of catabolic dehydrogenases by elevated moieties of ATP and ADP - implication for a new regulation mechanism in Lactococcus lactis

Cao, Rong LU ; Zeidan, Ahmad LU ; Rådström, Peter LU and van Niel, Ed LU (2010) In The FEBS Journal 277(8). p.1843-1852
Abstract
ATP and ADP inhibit, in varying degrees, several dehydrogenases of the

central carbon metabolism of Lactococcus lactis ATCC 19435 in vitro, i.e.

glyceraldehyde-3-phosphate dehydrogenase (GAPDH), lactate dehydrogenase

(LDH) and alcohol dehydrogenase (ADH). Here we demonstrate

mixed inhibition for GAPDH and competitive inhibition for LDH and

ADH by adenine nucleotides in single inhibition studies. The nonlinear

negative co-operativity was best modelled with Hill-type kinetics, showing

greater flexibility than the usual parabolic inhibition equation. Because

these natural inhibitors are present simultaneously in the cytoplasm, multiple

inhibition kinetics was... (More)
ATP and ADP inhibit, in varying degrees, several dehydrogenases of the

central carbon metabolism of Lactococcus lactis ATCC 19435 in vitro, i.e.

glyceraldehyde-3-phosphate dehydrogenase (GAPDH), lactate dehydrogenase

(LDH) and alcohol dehydrogenase (ADH). Here we demonstrate

mixed inhibition for GAPDH and competitive inhibition for LDH and

ADH by adenine nucleotides in single inhibition studies. The nonlinear

negative co-operativity was best modelled with Hill-type kinetics, showing

greater flexibility than the usual parabolic inhibition equation. Because

these natural inhibitors are present simultaneously in the cytoplasm, multiple

inhibition kinetics was determined for each dehydrogenase. For ADH

and LDH, the inhibitor combinations ATP plus NAD and ADP plus

NAD are indifferent to each other. Model discrimination suggested that

the weak allosteric inhibition of GAPDH had no relevance when multiple

inhibitors are present. Interestingly, with ADH and GAPDH the combination

of ATP and ADP exhibits lower dissociation constants than with

either inhibitor alone. Moreover, the concerted inhibition of ADH and

GAPDH, but not of LDH, shows synergy between the two nucleotides.

Similar kinetics, but without synergies, were found for horse liver and yeast

ADHs, indicating that dehydrogenases can be modulated by these nucleotides

in a nonlinear manner in many organisms. The action of an elevated

pool of ATP and ADP may effectively inactivate lactococcal ADH, but

not GAPDH and LDH, providing leverage for the observed metabolic shift

to homolactic acid formation in lactococcal resting cells on maltose. Therefore,

we interpret these results as a regulation mechanism contributing to

readjusting the flux of ATP production in L. lactis. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
ATP, ADP, Lactococcus lactis, multiple inhibition kinetics, dehydrogenase
in
The FEBS Journal
volume
277
issue
8
pages
1843 - 1852
publisher
Federation of European Neuroscience Societies and Blackwell Publishing Ltd
external identifiers
  • wos:000276168800006
  • scopus:77950496679
ISSN
1742-464X
DOI
10.1111/j.1742-4658.2010.07601.x
language
English
LU publication?
yes
id
e35b6038-c4c3-430a-a9b2-2a7fa5268579 (old id 1581335)
date added to LUP
2010-04-07 11:27:07
date last changed
2018-05-29 09:26:46
@article{e35b6038-c4c3-430a-a9b2-2a7fa5268579,
  abstract     = {ATP and ADP inhibit, in varying degrees, several dehydrogenases of the<br/><br>
central carbon metabolism of Lactococcus lactis ATCC 19435 in vitro, i.e.<br/><br>
glyceraldehyde-3-phosphate dehydrogenase (GAPDH), lactate dehydrogenase<br/><br>
(LDH) and alcohol dehydrogenase (ADH). Here we demonstrate<br/><br>
mixed inhibition for GAPDH and competitive inhibition for LDH and<br/><br>
ADH by adenine nucleotides in single inhibition studies. The nonlinear<br/><br>
negative co-operativity was best modelled with Hill-type kinetics, showing<br/><br>
greater flexibility than the usual parabolic inhibition equation. Because<br/><br>
these natural inhibitors are present simultaneously in the cytoplasm, multiple<br/><br>
inhibition kinetics was determined for each dehydrogenase. For ADH<br/><br>
and LDH, the inhibitor combinations ATP plus NAD and ADP plus<br/><br>
NAD are indifferent to each other. Model discrimination suggested that<br/><br>
the weak allosteric inhibition of GAPDH had no relevance when multiple<br/><br>
inhibitors are present. Interestingly, with ADH and GAPDH the combination<br/><br>
of ATP and ADP exhibits lower dissociation constants than with<br/><br>
either inhibitor alone. Moreover, the concerted inhibition of ADH and<br/><br>
GAPDH, but not of LDH, shows synergy between the two nucleotides.<br/><br>
Similar kinetics, but without synergies, were found for horse liver and yeast<br/><br>
ADHs, indicating that dehydrogenases can be modulated by these nucleotides<br/><br>
in a nonlinear manner in many organisms. The action of an elevated<br/><br>
pool of ATP and ADP may effectively inactivate lactococcal ADH, but<br/><br>
not GAPDH and LDH, providing leverage for the observed metabolic shift<br/><br>
to homolactic acid formation in lactococcal resting cells on maltose. Therefore,<br/><br>
we interpret these results as a regulation mechanism contributing to<br/><br>
readjusting the flux of ATP production in L. lactis.},
  author       = {Cao, Rong and Zeidan, Ahmad and Rådström, Peter and van Niel, Ed},
  issn         = {1742-464X},
  keyword      = {ATP,ADP,Lactococcus lactis,multiple inhibition kinetics,dehydrogenase},
  language     = {eng},
  number       = {8},
  pages        = {1843--1852},
  publisher    = {Federation of European Neuroscience Societies and Blackwell Publishing Ltd},
  series       = {The FEBS Journal},
  title        = {Inhibition kinetics of catabolic dehydrogenases by elevated moieties of ATP and ADP - implication for a new regulation mechanism in Lactococcus lactis},
  url          = {http://dx.doi.org/10.1111/j.1742-4658.2010.07601.x},
  volume       = {277},
  year         = {2010},
}