Functional expression of human cone pigments using recombinant baculovirus: Compatibility with histidine tagging and evidence for N-glycosylation

被引:28
作者
Vissers, PMAM [1 ]
DeGrip, WJ [1 ]
机构
[1] CATHOLIC UNIV NIJMEGEN,DEPT BIOCHEM,INST CELLULAR SIGNALLING,NL-6500 HB NIJMEGEN,NETHERLANDS
关键词
baculovirus; functional expression; human cone pigment; histidine tag; glycosylation; spectroscopy;
D O I
10.1016/0014-5793(96)01064-2
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Mammalian color vision is mediated by light-sensitive pigments in retinal cone cells, Biochemical studies on native mammalian cone visual pigments are seriously hampered by their low levels and instability. We describe a novel approach for their functional expression, employing the baculovirus system in combination with histidine tagging to allow future purification and structural analysis. The human red and green cone pigments are produced in relatively large amounts and can be detected by immunocytochemistry as sell as by immunoblotting. Histidine tagging has no significant effect on the absorbance maxima. The first evidence is presented that these pigments are N-glycosylated.
引用
收藏
页码:26 / 30
页数:5
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