| Literature DB >> 11532454 |
P J Werten1, L Hasler, J B Koenderink, C H Klaassen, W J de Grip, A Engel, P M Deen.
Abstract
The homotetrameric aquaporin-2 (AQP2) water channel is essential for the concentration of urine and of critical importance in diseases with water dysregulation, such as nephrogenic diabetes insipidus, congestive heart failure, liver cirrhosis and pre-eclampsia. The structure of human AQP2 is a prerequisite for understanding its function and for designing specific blockers. To obtain sufficient amounts of AQP2 for structural analyses, we have expressed recombinant his-tagged human AQP2 (HT-AQP2) in the baculovirus/insect cell system. Using the protocols outlined in this study, 0.5 mg of pure HT-AQP2 could be obtained per liter of bioreactor culture. HT-AQP2 had retained its homotetrameric structure and exhibited a single channel water permeability of 0.93+/-0.03x10(-13) cm3/s, similar to that of other AQPs. Thus, the baculovirus/insect cell system allows large-scale expression of functional recombinant human AQP2 that is suitable for structural studies.Entities:
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Year: 2001 PMID： 11532454 DOI： 10.1016/s0014-5793(01)02703-x
Source DB: PubMed Journal: FEBS Lett ISSN： 0014-5793 Impact factor: 4.124