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Isolation of a cDNA encoding a granule-bound 152-kilodalton starch-branching enzyme in wheat

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1104/pp.124.1.253
AuthorSearch for: 1; Search for: 1; Search for: 1; Search for: ; Search for: 1
Affiliation
  1. National Research Council Canada. NRC Plant Biotechnology Institute
FormatText, Article
Abstract
Screening of a wheat (Triticum aestivum) cDNA library for starch-branching enzyme I (SBEI) genes combined with 5′-rapid amplification of cDNA ends resulted in isolation of a 4,563-bp composite cDNA, Sbe1c. Based on sequence alignment to characterized SBEI cDNA clones isolated from plants, the SBEIc predicted from the cDNA sequence was produced with a transit peptide directing the polypeptide into plastids. Furthermore, the predicted mature form of SBEIc was much larger (152 kD) than previously characterized plant SBEI (80–100 kD) and contained a partial duplication of SBEI sequences. The first SBEI domain showed high amino acid similarity to a 74-kD wheat SBEI-like protein that is inactive as a branching enzyme when expressed in Escherichia coli. The second SBEI domain on SBEIc was identical in sequence to a functional 87-kD SBEI produced in the wheat endosperm. Immunoblot analysis of proteins produced in developing wheat kernels demonstrated that the 152-kD SBEIc was, in contrast to the 87- to 88-kD SBEI, preferentially associated with the starch granules. Proteins similar in size and recognized by wheat SBEI antibodies were also present inTriticum monococcum, Triticum tauschii, and Triticum turgidum subsp. durum.
Publication date
PublisherOxford Academic
American Society of Plant Biologists
In
  • Plant Physiology 124, no. 1 (1 September 2000): 253–262.
LanguageEnglish
Peer reviewedYes
NPARC number13053880
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Record identifier37c316b3-703a-43ba-a859-334b6b029516
Record created2009-11-25
Record modified2023-04-13
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