Sucrose synthase isoforms in cultured tobacco cells
(2004) In Plant Physiology and Biochemistry 42(4). p.299-306- Abstract
- The plant enzyme sucrose synthase (SuSy; EC 2.4.1.13) catalyzes the reversible conversion of sucrose and UDP into UDP-glucose (UDP-Glc) and fructose. The enzyme exists in different isoforms and is both located in the cytosol, membrane-bound and associated to the actin cytoskeleton. We here investigate sucrose synthase from tobacco (Nicotiana tabacum L.) BY-2 heterotrophic cell suspensions. Two different isoforms of sucrose synthase SuSy1 and SuSy2, could be purified from cytosolic extracts of these cells using a combination of poly(ethylene glycol) (PEG) precipitation, gel filtration, ion-exchange chromatography and affinity chromatography. They were clearly distinct. both with regard to the binding to the ion-exchange column and with... (More)
- The plant enzyme sucrose synthase (SuSy; EC 2.4.1.13) catalyzes the reversible conversion of sucrose and UDP into UDP-glucose (UDP-Glc) and fructose. The enzyme exists in different isoforms and is both located in the cytosol, membrane-bound and associated to the actin cytoskeleton. We here investigate sucrose synthase from tobacco (Nicotiana tabacum L.) BY-2 heterotrophic cell suspensions. Two different isoforms of sucrose synthase SuSy1 and SuSy2, could be purified from cytosolic extracts of these cells using a combination of poly(ethylene glycol) (PEG) precipitation, gel filtration, ion-exchange chromatography and affinity chromatography. They were clearly distinct. both with regard to the binding to the ion-exchange column and with regard to their kinetic and regulatory properties. SuSy 1, the more abundant species, showed lower V-max and K-m for sucrose and UDP compared to the less abundant SuSy2. The activity of SuSy2 in the breakdown direction was stimulated by 60% by actin, in contrast to that of SuSy 1, which showed a 17% inhibition. An indication of interaction between SuSy I and actin was obtained by partitioning in aqueous Dextran-PEG two-phase systems. Furthermore, fructose 2,6-bisphosphate (F26BP) at micromolar concentrations stimulated SuSy2 in the presence of actin while SuSy I was strongly inhibited by fructose. Possible roles of these two isoforms in the sucrose turnover in BY-2 cells are discussed. (C) 2004 Elsevier SAS. All rights reserved. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/138587
- author
- Matic, Sandra LU ; Åkerlund, Hans-Erik LU ; Everitt, Einar LU and Widell, Susanne LU
- organization
- publishing date
- 2004
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Two-phase partitioning, Tobacco, Sucrose synthase, Sucrolysis, 6-bisphosphate, Fructose 2, Actin, BY-2 cells
- in
- Plant Physiology and Biochemistry
- volume
- 42
- issue
- 4
- pages
- 299 - 306
- publisher
- Elsevier
- external identifiers
-
- pmid:15120114
- wos:000221421500005
- scopus:2342592684
- ISSN
- 1873-2690
- DOI
- 10.1016/j.plaphy.2004.01.009
- language
- English
- LU publication?
- yes
- id
- 5ea70c96-5160-44a9-ad1c-661690bbd1cb (old id 138587)
- date added to LUP
- 2016-04-01 15:48:03
- date last changed
- 2022-02-12 17:43:58
@article{5ea70c96-5160-44a9-ad1c-661690bbd1cb,
abstract = {{The plant enzyme sucrose synthase (SuSy; EC 2.4.1.13) catalyzes the reversible conversion of sucrose and UDP into UDP-glucose (UDP-Glc) and fructose. The enzyme exists in different isoforms and is both located in the cytosol, membrane-bound and associated to the actin cytoskeleton. We here investigate sucrose synthase from tobacco (Nicotiana tabacum L.) BY-2 heterotrophic cell suspensions. Two different isoforms of sucrose synthase SuSy1 and SuSy2, could be purified from cytosolic extracts of these cells using a combination of poly(ethylene glycol) (PEG) precipitation, gel filtration, ion-exchange chromatography and affinity chromatography. They were clearly distinct. both with regard to the binding to the ion-exchange column and with regard to their kinetic and regulatory properties. SuSy 1, the more abundant species, showed lower V-max and K-m for sucrose and UDP compared to the less abundant SuSy2. The activity of SuSy2 in the breakdown direction was stimulated by 60% by actin, in contrast to that of SuSy 1, which showed a 17% inhibition. An indication of interaction between SuSy I and actin was obtained by partitioning in aqueous Dextran-PEG two-phase systems. Furthermore, fructose 2,6-bisphosphate (F26BP) at micromolar concentrations stimulated SuSy2 in the presence of actin while SuSy I was strongly inhibited by fructose. Possible roles of these two isoforms in the sucrose turnover in BY-2 cells are discussed. (C) 2004 Elsevier SAS. All rights reserved.}},
author = {{Matic, Sandra and Åkerlund, Hans-Erik and Everitt, Einar and Widell, Susanne}},
issn = {{1873-2690}},
keywords = {{Two-phase partitioning; Tobacco; Sucrose synthase; Sucrolysis; 6-bisphosphate; Fructose 2; Actin; BY-2 cells}},
language = {{eng}},
number = {{4}},
pages = {{299--306}},
publisher = {{Elsevier}},
series = {{Plant Physiology and Biochemistry}},
title = {{Sucrose synthase isoforms in cultured tobacco cells}},
url = {{http://dx.doi.org/10.1016/j.plaphy.2004.01.009}},
doi = {{10.1016/j.plaphy.2004.01.009}},
volume = {{42}},
year = {{2004}},
}