Cinnamomin: separation, crystallization and preliminary X-ray diffraction study

被引:3
作者
Wang, T.
Zou, Y. -S.
Zhu, D. -W. [1 ]
Azzi, A. [1 ]
Liu, W. -Y.
Lin, S. -X. [1 ]
机构
[1] CHUQ, CHUL Med Ctr, Lab Oncol & Mol Endocrinol, Quebec City, PQ, Canada
关键词
cinnamomin; crystallization; FPLC; hydrophobic interaction chromatography; ribosome-inactivating protein; X-ray crystal structure;
D O I
10.1007/s00726-006-0490-z
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Cinnamomin from Cinnamonum camphora seeds, a type II ribosome-inactivating protein that interferes with protein biosynthesis in mammalian cells, can induce the apoptosis of carcinoma cells and be used as an insecticide. A rapid and improved method has been developed for the extraction and purification of cinnamomin from camphora seed. Purification of cinnamomin is achieved with two successive steps of hydrophobic interaction chromatography carried out on a fast protein liquid chromatography (FPLC) system. Crystals suitable for X-ray diffraction analysis were obtained by vapor diffusion method. A complete data set at 2.8 angstrom resolution has been collected. Data indexation and refinement indicate that the crystal is orthorhombic with space group P2(1)2(1)2(1) and unit cell dimensions a = 52.39 angstrom, b = 126.33 angstrom, c = 161.45 angstrom. There are two molecules per asymmetric unit. Initial phasing by molecular replacement method yielded a solution, which will contribute to the structure determination. A molecular model will further the understanding of the mechanism of cinnamomin function. The latter will be combined with bio-informatics to facilitate the medical and other applications of cinnamomin.
引用
收藏
页码:239 / 243
页数:5
相关论文
共 25 条
[11]   SIMULTANEOUS EXISTENCE OF 2 TYPES OF RIBOSOME-INACTIVATING PROTEINS IN THE SEEDS OF CINNAMONUM-CAMPHORA - CHARACTERIZATION OF THE ENZYMATIC-ACTIVITIES OF THESE CYTOTOXIC PROTEINS [J].
LING, J ;
LIU, WY ;
WANG, TP .
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY, 1995, 1252 (01) :15-22
[12]   Searching for hypothetical proteins: Theory and practice based upon original data and literature [J].
Lubec, G ;
Afjehi-Sadat, L ;
Yang, JW ;
John, JPP .
PROGRESS IN NEUROBIOLOGY, 2005, 77 (1-2) :90-127
[13]   Crystal structure of Himalayan mistletoe ribosome-inactivating protein reveals the presence of a natural inhibitor and a new functionally active sugar-binding site [J].
Mishra, V ;
Bilgrami, S ;
Sharma, RS ;
Kaur, P ;
Yadav, S ;
Krauspenhaar, R ;
Betzel, C ;
Voelter, W ;
Babu, CR ;
Singh, TP .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2005, 280 (21) :20712-20721
[14]  
MLSNA D, 1993, PROTEIN SCI, V2, P429
[15]   The finer things in X-ray diffraction data collection [J].
Pflugrath, JW .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1999, 55 :1718-1725
[16]   Purification and activity study of the A- and B-chains of cinnamomin, a type II ribosome-inactivating protein [J].
Pu, Z ;
Xie, L ;
Wang, ED ;
Liu, WY .
BIOLOGICAL CHEMISTRY, 1998, 379 (12) :1413-1418
[17]   JIP60, A METHYL JASMONATE-INDUCED RIBOSOME-INACTIVATING PROTEIN INVOLVED IN PLANT STRESS REACTIONS [J].
REINBOTHE, S ;
REINBOTHE, C ;
LEHMANN, J ;
BECKER, W ;
APEL, K ;
PARTHIER, B .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (15) :7012-7016
[18]   CRYSTAL-STRUCTURE OF ABRIN-A AT 2.14 ANGSTROM [J].
TAHIROV, TH ;
LU, TH ;
LIAW, YC ;
CHEN, YL ;
LIN, JY .
JOURNAL OF MOLECULAR BIOLOGY, 1995, 250 (03) :354-367
[19]   An approach to multi-copy search in molecular replacement [J].
Vagin, A ;
Teplyakov, A .
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2000, 56 :1622-1624
[20]   The lower cytotoxicity of cinnamomin (a type II RIP) is due to its B-chain [J].
Wang, Bao-Zhong ;
Zou, Wei-Guo ;
Liu, Wang-Yi ;
Liu, Xin-Yuan .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 2006, 451 (01) :91-96