Proteins that glow in green and blue.
Proteins that glow in green and blue.
复制标题
发出绿光和蓝光的蛋白质。
DOI：
10.1016/1074-5521(95)90011-x
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发表时间：
1995
影响因子：
--
通讯作者：
Bestor,TH
中科院分区：
文献类型：
--
作者：
Coxon,A;Bestor,TH
受到搅动时，水母 AC (IICMXYI victorid) 会从其触手之间发出绿光。这种绿光是两种密切相关的蛋白质的产物：一种称为水母发光蛋白的钙激活荧光素酶和绿色荧光蛋白 (GFP)。Aeyuorin 会产生蓝光；GFP 将此光的波长从蓝色 (hllldY 470 nm) 更改为绿色 (hlllaX 509 nm)[l]。两种蛋白质之间的能量转移被认为是通过非辐射机制发生的 121。值得注意的是，GFP 将蓝光转换为绿光，而不需要辅助因子或外源底物；荧光团由蛋白质内明显前所未有的环状三肽组成，该三肽似乎是由一种新的自催化机制形成的[3， 4]。 GFP现已在从大肠杆菌到哺乳动物细胞的各种生物体中表达，并且已发现在所测试的每种细胞类型中都具有强烈的荧光。野生型GFP是一种分子量为27 kDa的单体蛋白。它的激发最大值为 395 nm，次要最大值为 475 nm；发光峰值在 509 nm [l]。该荧光团的独特之处在于它是通过三肽环化和随后环化酪氨酰侧链 151 的氧化而衍生的； Heim等人提出的反应机理[5]是
Upon agitation, the jellyfish AC (IICMXYI victorid emits green light from between its tentacles. This green light is the product of two closely-associated proteins: a calciumactivated luciferase known as aequorin, and green fluorescent protein (GFP). Aeyuorin generates blue light; GFP alters the wavelength of this light from blue (hllldY 470 nm) to green (hlllaX 509 nm)[l]. The transfer of energy between the two proteins is thought to take place by a non-radiative mechanism 121. Remarkably, GFP converts the blue light to green without the need for cofactors or exogenous substrates; the fluorophore consists of an apparently unprecedented cyclic tripeptide within the protein that appears to be formed by a novel autocatalytic mechanism[3, 4]. GFP has now been expressed in organisms ranging from Escherichia coli to mammalian cells, and has been found to be strongly fluorescent in every cell type tested.Wild-type GFP is a monomeric protein with a molecular weight of 27 kDa. It has an excitation maximum of 395 nm with a secondary maxitnum at 475 nm; light emission peaks at 509 nm [l]. The fluorophore is unique in that it is derived by cyclization of a tripeptide and subsequent oxidation of the cyclized tyrosyl side chain 151; a reaction mechanism proposed by Heim et al.[5] is