Identification of the major starch synthase in the soluble fraction of potato tubers.
The major isoform of starch synthase from the soluble fraction of developing potato tubers has been purified and used to prepare an antibody and isolate a cDNA. The protein is 140 kD, and it is distinctly different in predicted primary amino acid sequence from other isoforms of the enzyme thus far described. Immunoinhibition and immunoblotting experiments and analysis of tubers in which activity of the isoform was reduced through expression of antisense mRNA revealed that the isoform accounts for approximately 80% of the activity in the soluble fraction of the tuber and that it is also bound to starch granules. Severe reductions in activity had no discernible effect on starch content or amylose-to-amylopectin ratio of starch in tubers. However, they caused a profound change in the morphology of starch granules, indicative of important underlying changes in the structure of starch polymers within the granule.
Marshall
J
J
John Innes Centre, Norwich, United Kingdom.
Sidebottom
C
C
Debet
M
M
Martin
C
C
Smith
A M
AM
Edwards
A
A
D000595:Amino Acid Sequence; D000906:Antibodies; D001419:Bacteria / Q000201:enzymology; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology; D018076:DNA, Complementary; D015870:Gene Expression; D007700:Kinetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D018517:Plant Roots; D016372:RNA, Antisense; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D011198:Solanum tuberosum / Q000201:enzymology*; D013214:Starch Synthase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*
D000906:Antibodies; D018076:DNA, Complementary; D016372:RNA, Antisense; D012333:RNA, Messenger; D011994:Recombinant Proteins; D013214:Starch Synthase