Biochemical and molecular characterization and expression of the 11S-type storage protein from Coffea arabica endosperm

✍ Scribed by W.John Rogers; Guy Bézard; Alain Deshayes; Inge Meyer; Vincent Pétiard; Pierre Marraccini

• Publisher: Elsevier Science
• Year: 1999
• Tongue: English
• Weight: 504 KB
• Volume: 37
• Category: Article
• ISSN: 0981-9428
• DOI: 10.1016/s0981-9428(99)80024-2

✦ Synopsis

In order to define better the endosperm protein content of commercial coffee species Coffea arabica (Arabica) and C. canephora (Robusta), the principal storage protein of coffee grains has been analysed by 2-dimensional electrophoresis (2DE) and amino acid microsequencing. The most abundant polypeptide spots observed on mature coffee grain 2DE profiles were found to be subunits of the same protein, which exists as multiple isoforms with varying pIs. Strong sequence similarity was found to the 11S family of plant storage proteins. The structure is typical of the 11S type, which occurs as a precursor of 55 kDa, and is observed under denaturing and reducing conditions on 2DE profiles in the form of cleavage products at approximately 20 kDa (b arms) and 32 kDa (a arms). Differences between Arabica and Robusta 2DE profiles indicate a secondary 11S protein family in some varieties of the latter. The existence of multiple pI forms may indicate that a multigene family encodes for these proteins. We estimate that the protein accounts for approximately 45 % of total grain protein. A cloned full-length cDNA of 1 706 bp coding for one of the isoforms is described and discussed in relation to other coffee storage protein sequences. © Elsevier, Paris Coffea arabica / Coffea canephora / 11S storage protein / endosperm / full-length cDNA / storage protein / two dimensional electrophoresis 2DE, immobilized pH gradient 2-dimensional electrophoresis / CHAPS, 3-[3(cholamidopropyl)-dimethylammonio]-1propanesulfonate / DTT, dithiothreitol / SDS-PAGE, SDS polyacrylamide gel electrophoresis / TCA, trichloroacetic acid / WAF, weeks after flowering