Primary structure and allergenic activity of trypsin inhibitors from the seeds of buckwheat (Fagopyrum esculentum Moench)

FEBS Letters

Kimura, Makoto¹  Abe, Kei¹  Yamasaki, Nobuyuki¹  Urisu, Atsuo²  Park, Sung-Soo¹ 

[1] Laboratory of Biochemistry, Faculty of Agriculture, Kyushu University, Fukuoka 812, Japan;Department of Pediatrics, Fujita Health University, School of Medicine, Dengakugakubo, Kutsukake-cho, Toyoake, Aichi 470-11, Japan

关键词: Buckwheat trypsin inhibitor;    Fagopyrum esculentum Moench;    Amino acid sequence;    Allergenic protein;    MALDI-TOF-MS;    matrix-assisted laser desorption and ionization time-of-flight mass spectrometry;    PE;    pyridylethyl;    RAST;    radioallergosorbent test;    RP-HPLC;    reverse-phase high-performance liquid chromatography;    TFA;    trifluoroacetic acid;    SCB;    sodium carbonate buffer;    PBS;    phosphate-buffered saline;   

DOI  :  10.1016/S0014-5793(96)01367-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The complete amino acid sequences of two trypsin inhibitors BWI-2a and BWI-2b from the seeds of buckwheat (Fagopyrum esculentum Moench) were determined. BWI-2b consists of 51 amino acid residues containing two disulfide bonds. BWI-2a shares all amino acids with BWI-2b except for the C-terminal tripeptide: BWI-2a lacks Glu-Gly-Asn and ends with the Asp residue, making a total of 48 residues in the chain. The two disulfide bonds connect Cys¹¹ to Cys³² and Cys¹⁵ to Cys²⁸. BWI-2b shows no relatedness to the other buckwheat trypsin inhibitor reported [Belozersky et al. (1995) FEBS Lett. 371, 264–266]. Sequence comparison of BWI-2b with those of the other proteins included in PIR showed that BWI-2b is significantly homologous to the N-terminal region of storage proteins classified in the vicilin family. Furthermore, the allergenic activity of BWI-2b and the other buckwheat trypsin inhibitor BWI-1 was examined using the radioallergosorbent test. The result indicated that both inhibitors BWI-2b and BWI-1 have IgE binding activity, albeit to a low extent, suggesting that they might be minor allergenic proteins in buckwheat seeds.

【 授权许可】

Unknown