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Functional and structural characterization of domain truncated violaxanthin de-epoxidase

Hallin, Erik Ingmar LU ; Guo, Kuo LU and Åkerlund, Hans Erik LU (2016) In Physiologia Plantarum 157(4). p.414-421
Abstract

Photosynthetic organisms need protection against excessive light. By using non-photochemical quenching, where the excess light is converted into heat, the organism can survive at higher light intensities. This process is partly initiated by the formation of zeaxanthin, which is achieved by the de-epoxidation of violaxanthin and antheraxanthin to zeaxanthin. This reaction is catalyzed by violaxanthin de-epoxidase (VDE). VDE consists of three domains of which the central lipocalin-like domain has been the most characterized. By truncating the domains surrounding the lipocalin-like domain, we show that VDE activity is possible without the C-terminal domain but not without the N-terminal domain. The N-terminal domain shows no VDE activity... (More)

Photosynthetic organisms need protection against excessive light. By using non-photochemical quenching, where the excess light is converted into heat, the organism can survive at higher light intensities. This process is partly initiated by the formation of zeaxanthin, which is achieved by the de-epoxidation of violaxanthin and antheraxanthin to zeaxanthin. This reaction is catalyzed by violaxanthin de-epoxidase (VDE). VDE consists of three domains of which the central lipocalin-like domain has been the most characterized. By truncating the domains surrounding the lipocalin-like domain, we show that VDE activity is possible without the C-terminal domain but not without the N-terminal domain. The N-terminal domain shows no VDE activity by itself but when separately expressed domains are mixed, VDE activity is possible. This shows that these domains can be folded separately and could therefore be studied separately. An increase of the hydrodynamic radius of wild-type VDE was observed when pH was lowered toward the pH required for activity, consistent with a pH-dependent oligomerization. The C-terminally truncated VDE did not show such an oligomerization, was relatively more active at higher pH but did not alter the KM for ascorbate. Circular dichroism measurements revealed the presence of α-helical structure in both the N- and C-terminal domains. By measuring the initial formation of the product, VDE was found to convert a large number of violaxanthin molecules to antheraxanthin before producing any zeaxanthin, favoring a model where violaxanthin is bound non-symmetrically in VDE.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Physiologia Plantarum
volume
157
issue
4
pages
8 pages
publisher
Wiley Online Library
external identifiers
  • pmid:26864799
  • scopus:84962832835
  • scopus:84978817827
  • wos:000380063100002
ISSN
0031-9317
DOI
10.1111/ppl.12428
language
English
LU publication?
yes
id
0be4bede-a788-43a2-a2b5-2407817df2a9 (old id 8825835)
date added to LUP
2016-03-07 16:09:10
date last changed
2017-01-01 05:47:16
@article{0be4bede-a788-43a2-a2b5-2407817df2a9,
  abstract     = {<p>Photosynthetic organisms need protection against excessive light. By using non-photochemical quenching, where the excess light is converted into heat, the organism can survive at higher light intensities. This process is partly initiated by the formation of zeaxanthin, which is achieved by the de-epoxidation of violaxanthin and antheraxanthin to zeaxanthin. This reaction is catalyzed by violaxanthin de-epoxidase (VDE). VDE consists of three domains of which the central lipocalin-like domain has been the most characterized. By truncating the domains surrounding the lipocalin-like domain, we show that VDE activity is possible without the C-terminal domain but not without the N-terminal domain. The N-terminal domain shows no VDE activity by itself but when separately expressed domains are mixed, VDE activity is possible. This shows that these domains can be folded separately and could therefore be studied separately. An increase of the hydrodynamic radius of wild-type VDE was observed when pH was lowered toward the pH required for activity, consistent with a pH-dependent oligomerization. The C-terminally truncated VDE did not show such an oligomerization, was relatively more active at higher pH but did not alter the K<sub>M</sub> for ascorbate. Circular dichroism measurements revealed the presence of α-helical structure in both the N- and C-terminal domains. By measuring the initial formation of the product, VDE was found to convert a large number of violaxanthin molecules to antheraxanthin before producing any zeaxanthin, favoring a model where violaxanthin is bound non-symmetrically in VDE.</p>},
  author       = {Hallin, Erik Ingmar and Guo, Kuo and Åkerlund, Hans Erik},
  issn         = {0031-9317},
  language     = {eng},
  month        = {08},
  number       = {4},
  pages        = {414--421},
  publisher    = {Wiley Online Library},
  series       = {Physiologia Plantarum},
  title        = {Functional and structural characterization of domain truncated violaxanthin de-epoxidase},
  url          = {http://dx.doi.org/10.1111/ppl.12428},
  volume       = {157},
  year         = {2016},
}