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Reconstitution of active sucrose transport in plant proteoliposomes
Author(s) -
Li Ze-Sheng,
Gallet Olivier,
Gaillard Cécile,
Lemoine Rémi,
Delrot Serge
Publication year - 1991
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(91)80954-2
Subject(s) - sucrose , chemistry , chromatography , valine , size exclusion chromatography , biochemistry , n ethylmaleimide , membrane , amino acid , enzyme
The proteins of purified plasma membranes from sugar beet ( Beta vulgaris L.) leaf were solubilized and separated on a size exclusion column. The fractions eluted from the column were monitored by ELISA with antibodies directed to a putative sucrose carrier protein. The peak most reactive in ELISA was approximately 120 kDa, and yielded a 40 kDa peak after denaturation by SDS. The 120‐kDa peak was recovered and used for reconstitution experiments with asolectin. Upon imposition of an artificial pH gradient and electrical gradient, the obtained proteoliposomes exhibited active transport of sucrose, but not of valine. The active transport of sucrose was inhibited by N ‐ethylmaleimide and HgCl 2 .