Mass spectrometry and structural characterization of 2S albumin isoforms from Brazil nuts (Bertholletia excelsa).
Proteomic approaches have been used to characterise the main 2S albumin isoforms from Brazil nuts (Bertholletia excelsa). Whilst most isoforms ( approximately 10 discrete protein species) exhibited molecular masses of around 12 kDa with a high amino acid sequence homology, important charge heterogeneity was found, with pIs varying between 4.6 and 6.6, with one >or=7.0. Proteomic analysis showed that these corresponded to a total of six National Center for Biotechnology Information (NCBI) accessions and that three isoforms had been purified to homogeneity corresponding to gi/384327, 112754 and 99609. The latter sequence corresponds to an isoform, previously only identified at the nucleotide sequence level, had a slightly higher molecular weight (13.4 kDa), and with noticeable differences in the primary structure. Proteins corresponding to six different NCBI accessions were identified, the heterogeneity of which had been increased by posttranslational processing. Evidence was found of cyclization of the N-terminal glutamine residue in two isoforms, together with ragged C-termini, indicative of carboxypeptidase activity within the vacuole following posttranslational processing. No evidence of glycosylation was found. Circular dichroism (CD) and Fourier transform-infrared (FT-IR) spectroscopy indicated all the studied isoforms were predominantly alpha-helical in nature, but that the Mr 13400 species was structurally distinct, with a higher proportion of alpha-helical structure.
Moreno
F Javier
FJ
Institute of Food Research, Norwich Research Park, Colney Lane, Norwich NR4 7UA, UK. javier.moreno@bbsrc.ac.uk
Jenkins
John A
JA
Mellon
Fred A
FA
Rigby
Neil M
NM
Robertson
James A
JA
Wellner
Nikolaus
N
Clare Mills
E N
EN
D055516:2S Albumins, Plant; D000418:Albumins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D031383:Bertholletia / Q000737:chemistry* / Q000235:genetics; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D014644:Genetic Variation; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics; D011498:Protein Precursors / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D017433:Protein Structure, Secondary; D017550:Spectroscopy, Fourier Transform Infrared
D055516:2S Albumins, Plant; D000418:Albumins; D052179:Antigens, Plant; D020033:Protein Isoforms; D011498:Protein Precursors; C103364:2S albumin, brazil nut