Recombinant human thyroid peroxidase expressed in insect cells is soluble at high concentrations and forms diffracting crystals

被引:25
作者
Hendry, E
Taylor, G [1 ]
Ziemnicka, K
Jones, FG
Furmaniak, J
Smith, BR
机构
[1] Univ Bath, Dept Biol & Biochem, Bath BA2 7AY, Avon, England
[2] RSR Ltd, FIRS Labs, Cardiff CF4 5DU, S Glam, Wales
[3] Cardiff Univ, Dept Med, Cardiff CF4 4XN, S Glam, Wales
基金
加拿大自然科学与工程研究理事会;
关键词
D O I
10.1677/joe.0.160R013
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Human thyroid peroxidase (TPO), the key enzyme in thyroid hormone synthesis, can be produced in active form in the High Five insect cell line and when purified from the cell culture medium is soluble at concentrations of up to 18 mg/ml. This contrasts to a recent report in which human TPO produced in insect cells was found to be insoluble at high concentrations. Our concentrated TPO grows trigonal trapezohedral crystals Of UP to 0.5 mm in length in a vapour diffusion apparatus using polyethelene glycol as a precipitant. The crystals diffract X-rays to 6 Angstrom resolution and the diffraction data from the crystals have been analysed giving unit cell, dimensions. A potential molecular replacement solution has been identified using myeloperoxidase (MPO) as a phasing model.
引用
收藏
页码:R13 / R15
页数:3
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