Purification and characterization of the antitumor protein from Chinese tartary buckwheat (Fagopyrum tataricum Gaertn.) water-soluble extracts.
A novel antitumor protein, coded as TBWSP31, was isolated from tartary buckwheat water-soluble extracts and purified by DEAE-Sepharose Fast Flow anion exchange, Sephadex G-100 gel filtration, and Sephacryl S-200 gel filtration column chromatography. TBWSP31 was identified to a homogeneous fraction by native PAGE. The antitumor effect of TBWSP31 against human mammary cancer cell Bcap37 was measured by an MTT assay. TBWSP31 showed higher antitumor activity, and time- and concentration-dependent effects were observed. SDS-PAGE analysis showed that TBWSP31 is composed of a single polypeptide with an approximate molecular weight of 57 kDa. TBWSP31 was rich in Glx (Gln+Glu), Arg, and Asx (Asp+Asn) according to amino acid analysis. Secondary structural analysis by CD spectroscopy revealed that TBWSP31 has the following: alpha-helix (33.9%), beta-sheet (22.8%), beta-turn (11.3%), and random coil (32.0%).
Guo
Xiaona
X
State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi-214122 Jiangsu Province, People's Republic of China. xiaonaguo@jiangnan.edu.cn
Zhu
Kexue
K
Zhang
Hui
H
Yao
Huiyuan
H
D000596:Amino Acids / Q000032:analysis; D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification* / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor; D019612:Fagopyrum / Q000737:chemistry*; D006801:Humans; D010936:Plant Extracts / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D000091402:Prohibitins; D012995:Solubility; D014867:Water
D000596:Amino Acids; D000972:Antineoplastic Agents, Phytogenic; C000717331:PHB2 protein, human; D010936:Plant Extracts; D010940:Plant Proteins; D000091402:Prohibitins; D014867:Water