Partial amino acid sequence of gamma-46 gliadin.
We have determined the partial amino acid sequence (207 amino acids) of gamma-46 gliadin isolated from wheat cultivar Hardi. The molecular mass of the protein (Mr) estimated by electrospray mass spectrometry is 35191.3. The number of cysteine residues in gamma-46 gliadin was determined as a mass difference of the protein before and after reduction and alkylation with 4-vinylpyridine. It was shown that the protein has no free SH-groups, and all cysteine residues are involved in the formation of four disulfide bonds. The partial structure of gamma-46 gliadin was determined by N-terminal sequencing and sequencing of tryptic and chymotryptic peptides. The tryptic peptides were obtained by enzymatic hydrolysis of the protein, which was preliminarily reduced and immobilized at free SH-groups on thiopropyl-Sepharose 6B. The chymotryptic peptides were isolated by limited digestion of the native protein. The positions of cysteine residues, as well as surrounding amino acid sequences, are conserved in gamma-46 gliadin; this is typical of gliadins.
Egorov
T A
TA
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 117984, Russia. egorov@imb.imb.ac.ru.
Odintsova
T I
TI
Musolyamov
A K
AK
Barbashov
S F
SF
Pustobaev
V N
VN
Andersen
J
J
Roepstorff
P
P
Popineau
Y
Y
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D002918:Chymotrypsin; D003545:Cysteine / Q000032:analysis; D005903:Gliadin / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D006868:Hydrolysis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D014908:Triticum / Q000737:chemistry / Q000235:genetics
D010446:Peptide Fragments; D005903:Gliadin; D002918:Chymotrypsin; D003545:Cysteine