Expression and structural characterization of peripherin/RDS, a membrane protein implicated in photoreceptor outer segment morphology
Expression and structural characterization of peripherin/RDS, a membrane protein implicated in photoreceptor outer segment morphology
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DOI：
10.1007/s00249-009-0553-7
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发表时间：
2010-03
期刊：
影响因子：
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通讯作者：
W. Vos;Sebastian Vaughan;P. Lall;J. McCaffrey;Monika Wysocka-Kapcińska;J. Findlay
中科院分区：
文献类型：
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作者：
W. Vos;Sebastian Vaughan;P. Lall;J. McCaffrey;Monika Wysocka-Kapcińska;J. Findlay
Peripherin/RDS is a member of the tetraspanin family of integral membrane proteins and plays a major role in the morphology of photoreceptor outer segments. Peripherin/RDS has a long extracellular loop (hereafter referred to as the LEL domain), which is vital for its function. Point mutations in the LEL domain often lead to impaired photoreceptor formation and function, making peripherin/RDS an important drug target. Being a eukaryotic membrane protein, acquiring sufficient peripherin/RDS for biophysical characterisation represents a significant challenge. Here, we describe the expression and characterisation of peripherin/RDS inDrosophila melangolasterSchneider (S2) insect cells and in the methylotrophic yeastPichia pastoris. The wild-type peripherin/RDS and theretinitis pigmentosacausing P216L mutant from S2 cells are characterised using circular dichroism (CD) spectroscopy. The structure of peripherin/RDS and of a pathogenic mutant is assessed spectroscopically for the first time. These findings are evaluated in relation to a three-dimensional model of the functionally important LEL domain obtained by protein threading.
