TY  - JOUR
DA  - 2014//
VL  - 9
IS  - 7
SP  - e102364
AU  - Brust, Henrike
AU  - Lehmann, Tanja
AU  - D'hulst, Christophe
AU  - Fettke, Joerg
TI  - Analysis of the functional interaction of Arabidopsis starch synthase and branching enzyme isoforms reveals that the cooperative action of SSI and BEs results in glucans with polymodal chain length distribution similar to amylopectin
DO  - 10.1371/journal.pone.0102364
JA  - PLoS ONE
LA  - Anglais
UR  - http://hdl.handle.net/20.500.12210/18355
SN  - 1932-6203
AB  - Starch synthase (SS) and branching enzyme (BE) establish the two glycosidic linkages existing in starch. Both enzymes exist as several isoforms. Enzymes derived from several species were studied extensively both in vivo and in vitro over the last years, however, analyses of a functional interaction of SS and BE isoforms are missing so far. Here, we present data from in vitro studies including both interaction of leaf derived and heterologously expressed SS and BE isoforms. We found that SSI activity in native PAGE without addition of glucans was dependent on at least one of the two BE isoforms active in Arabidopsis leaves. This interaction is most likely not based on a physical association of the enzymes, as demonstrated by immunodetection and native PAGE mobility analysis of SSI, BE2, and BE3. The glucans formed by the action of SSI/BEs were analysed using leaf protein extracts from wild type and be single mutants (Atbe2 and Atbe3 mutant lines) and by different combinations of recombinant proteins. Chain length distribution (CLD) patterns of the formed glucans were irrespective of SSI and BE isoforms origin and still independent of assay conditions. Furthermore, we show that all SS isoforms (SSI-SSIV) were able to interact with BEs and form branched glucans. However, only SSI/BEs generated a polymodal distribution of glucans which was similar to CLD pattern detected in amylopectin of Arabidopsis leaf starch. We discuss the impact of the SSI/BEs interplay for the CLD pattern of amylopectin.
KW  - Arabidopsis Proteins
KW  - Signal Transduction
KW  - Starch Synthase
KW  - Isoenzymes
KW  - Arabidopsis
KW  - Starch
KW  - Amylopectin
KW  - Plant Leaves
KW  - Gene Expression Regulation, Plant
KW  - 1,4-alpha-Glucan Branching Enzyme
KW  - Recombinant Proteins
JO  - PLoS One
ER  - 
