Introduction of a rod pigment aromatic cluster does not improve the structural stability of the human green cone pigment.

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Publication year
2007Source
Journal of Structural Biology, 159, 2, (2007), pp. 222-7ISSN
Publication type
Article / Letter to editor

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Organization
Biochemistry (UMC)
Journal title
Journal of Structural Biology
Volume
vol. 159
Issue
iss. 2
Page start
p. 222
Page end
p. 7
Subject
DCN 1: Perception and Action; IGMD 5: Health aging / healthy living; NCMLS 1: Immunity, infection and tissue repair; UMCN 1.4: Immunotherapy, gene therapy and transplantationAbstract
In the course of our studies on the structure/function relationship of visual pigments, we have expressed the human green cone pigment in the baculovirus/insect cell expression system. Purification of the human green cone pigment, however, has so far proven to be severely hampered by the low thermal stability of this receptor in a solubilized state. In order to overcome this problem, we tested a variety of chemical compounds that have been described to improve protein stability in various applications. The presence of glycerol, sucrose, trehalose and lipids during extraction improved the thermal stability of the recombinant green cone pigment up to twofold. We also analyzed the effect of mutation of residues Met208, Cys212 and Cys273 into Phe in all combinations. These mutants were designed in an attempt to increase the thermal stability by replacing weakly interacting side chains in the green pigment with their counterparts in rhodopsin with strong aromatic stacking interaction. All mutants produced wild-type levels of functional pigment, but none showed an increase in thermal stability.
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- Academic publications [227942]
- Electronic publications [107434]
- Faculty of Medical Sciences [86237]
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