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Titolo:
Location of disulfide bonds in mature alpha-L-fucosidase from pea
Autore:
Codina, A; Vilaseca, M; Tarrago, T; Fernandez, I; Ludevid, D; Giralt, E;
Indirizzi:
Univ Barcelona, Dept Organ Chem, E-08028 Barcelona, Spain Univ Barcelona Barcelona Spain E-08028 an Chem, E-08028 Barcelona, Spain Univ Barcelona, Mass Spectrometry Serv, Div 3, E-08028 Barcelona, Spain Univ Barcelona Barcelona Spain E-08028 , Div 3, E-08028 Barcelona, Spain CSIC, Cid, Dept Mol Genet, ES-08034 Barcelona, Spain CSIC Barcelona Spain ES-08034 Dept Mol Genet, ES-08034 Barcelona, Spain
Titolo Testata:
JOURNAL OF PEPTIDE SCIENCE
fascicolo: 6, volume: 7, anno: 2001,
pagine: 305 - 315
SICI:
1075-2617(200106)7:6<305:LODBIM>2.0.ZU;2-Z
Fonte:
ISI
Lingua:
ENG
Soggetto:
FLIGHT MASS-SPECTROMETRY; PROTEIN IDENTIFICATION; CYSTEINE RESIDUES; PEPTIDE; PURIFICATION; MALDI;
Keywords:
acrylamide; alpha-L-fucosidase; disulfide bonds; free cysteines; Fuc-9; MALDI-TOF mass spectrometry; NTCB; tryptic digestion;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
14
Recensione:
Indirizzi per estratti:
Indirizzo: Giralt, E Univ Barcelona, Dept Organ Chem, Marti & Franques 1, E-08028 Barcelona, Spain Univ Barcelona Marti & Franques 1 Barcelona Spain E-08028 Spain
Citazione:
A. Codina et al., "Location of disulfide bonds in mature alpha-L-fucosidase from pea", J PEPT SCI, 7(6), 2001, pp. 305-315

Abstract

Fuc-9 is the mature form of a vacuolar alpha -L-fucosidase enzyme which seems to play an important role In plant growth regulation. Fuc-9 is a 202-residue protein containing five Cys residues located at positions 64, 109, 127, 162 and 169. In this study. the disulfide structure of Fuc-9 was determined by MALDI-TOF mass spectrometry (MS), with minimal clean-up of the samples and at a nanomolar scale. Two strategies, based on a specific chemical cleavage (with 2-nitro-5-thiocyanobenzoic acid and alkaline conditions) at the Cys residues and modification of Cys residues by acrylamide/deuterium labeled acrylamide alkylation, were used. Using these methods. the disulfide pairings CysB4-Cys109 and Cys162-Cys169 could be established. The advantages and limitations of our experimental approach are discussed. Copyright (C)2001 European Peptide Society and John Wiley & Sons, Ltd.

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Documento generato il 29/09/20 alle ore 10:19:32