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GrainGenes Reference Report: JCS-34-85

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Reference	JCS-34-85
Title	Albumin and globulin proteins of wheat flour: Immunological and N-terminal sequence characterisation
Journal	Journal of Cereal Science
Year	2001
Volume	34
Pages	85-103
Author	Singh J Blundell M Tanner G Skerritt JH
Abstract	Summary: Several non-storage proteins are encoded by genes on individual wheat chromosomes and these have been used as genome-specific markers. In this study, a library of monoclonal antibodies with differing specificities to water- and salt-soluble proteins has been developed in order to obtain markers for different wheat chromosomes. Two antibodies, BCSAU 9D1 and JGM 1B4 were found to bind polypeptides encoded by chromosomes 3D and 4D respectively. Other antibodies bound to small numbers of polypeptides encoded by more than one chromosome, such as 5A, 7D, 5B and 5D. The proteins recognised by some of the antibodies were isolated by immunoaffinity chromatography, and one protein was identified as an alpha-amylase inhibitor by N-terminal sequence characterisation. In addition, 16 water-soluble and eight salt-soluble proteins were isolated using SDS-PAGE, isoelectric focusing, two-dimensional electrophoresis and reversed-phase high performance liquid chromatography, with the main objective being to confirm the identity of particular proteins, especially those which were able to be assigned to particular chromosomes. The N-terminal sequencing characterisation identified 19 proteins, whereas four proteins were found to be blocked and one did not match with any proteins in the database. Most of the water-soluble proteins belonged to a family of alpha-amylase inhibitors. One protein, assigned to chromosome 4BS, was homologous to serine carboxypeptidase III. N-terminal sequences of some of salt-soluble proteins matched with internal sequences of barley embryo globulin. Other proteins were identified as lipid transfer protein, peroxidase BP-1 precursor and histone H4 proteins. The protein sequences could also potentially be used for making antibody or DNA probes for use in selection in breeding programmes
Keyword	[ Hide all but 1 of 58 ] 2-dimensional electrophoresis 5a addition albumin alpha-amylase alpha-amylase inhibitor antibody barley barley embryo breeding breeding programme chromatography chromosomal location chromosome database dna dna probes electrophoresis embryo endosperm family flour food gene globulins h4 high performance liquid chromatography histone inhibitor library lipid lipid transfer protein liquid chromatography marker monoclonal antibodies n-terminal sequence n-terminal sequencing performance peroxidase polypeptide precursor probes protein protein sequences reversed-phase sds page selection sequence sequencing serine serine-carboxypeptidase soluble proteins specificity transfer water wheat chromosomes wheat flour wheat grain

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