Proteomic identification of alpha-amylase isoforms encoded by RAmy3B/3C from germinating rice [Oryza sativa] seeds

We isolated and identified 10 α-amylase isoforms by using β-cyclodextrin Sepharose affinity column chromatography and two-dimensional polyacrylamide gel electrophoresis from germinating rice (Oryza sativa L.) seeds. Immunoblots with anti-α-amylase I-1 and II-4 antibodies indicated that 8 isoforms in...

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Bibliographic Details
Published in:Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2004-01, Vol.68 (1), p.112-118
Main Authors: Nanjo, Y. (Niigata Univ. (Japan)), Asatuma, S, Itoh, K, Hori, H, Mitsui, T
Format: Article
Language:English
Subjects:
ALPHA AMYLASE
alpha-Amylases - genetics
alpha-Amylases - isolation & purification
alpha-Amylases - metabolism
Amino Acid Sequence
Biological and medical sciences
Chromatography, Affinity - methods
DNA FINGERPRINTING
ELECTROPHORESIS
Fundamental and applied biological sciences. Psychology
GENES
Genes, Plant
germinating rice seed
GERMINATION
IDENTIFICATION
IMMUNOBLOTTING
Isoenzymes
Molecular Sequence Data
Oryza - enzymology
Oryza - genetics
ORYZA SATIVA
PEPTIDES
POLYMORPHISM
Polymorphism, Genetic
proteomics
Proteomics - methods
RAmy3B
RAmy3C
Seeds - enzymology
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods
Temperature
α-amylase
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Summary:We isolated and identified 10 α-amylase isoforms by using β-cyclodextrin Sepharose affinity column chromatography and two-dimensional polyacrylamide gel electrophoresis from germinating rice (Oryza sativa L.) seeds. Immunoblots with anti-α-amylase I-1 and II-4 antibodies indicated that 8 isoforms in 10 are distinguishable from α-amylase I-1 and II-4. Peptide mass fingerprinting analysis showed that there exist novel isoforms encoded by RAmy3B and RAmy3C genes. The optimum temperature for enzyme reaction of the RAmy3B and RAmy3C coding isoforms resembled that of α-amylase isoform II-4 (RAmy3D). Furthermore, complex protein polymorphism resulted from a single α-amylase gene was found to occur not only in RAmy3D, but also in RAmy3B.
ISSN:0916-8451
1347-6947
DOI:10.1271/bbb.68.112