Cloning, sequence analysis, and expression of active Phrixothrix railroad-worms luciferases:: Relationship between bioluminescence spectra and primary structures
Cloning, sequence analysis, and expression of active Phrixothrix railroad-worms luciferases:: Relationship between bioluminescence spectra and primary structures
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DOI：
10.1021/bi9900830
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发表时间：
1999-06-29
期刊：
影响因子：
2.9
通讯作者：
Ohmiya, Y
中科院分区：
文献类型：
--
作者：
Viviani, VR;Bechara, EJH;Ohmiya, Y
Phrixothrix railroad-worms发出黄绿色光通过11对侧灯笼沿着身体和红色光通过两个头部灯笼。克隆了发绿色光（λ（max）= 542 nm）的Vivianii Phrixothrix的侧灯笼荧光素酶的cDNA和在发光甲虫中发出最红移的生物发光（λ（max）= 628 nm）的P.hirtus的头灯笼的cDNA。分离发射绿色（Pv（GR）：λ（max）= 549 nm）和红色（Ph-RE：λ（max）= 622 nm）生物发光的阳性克隆。Pv（GR）和Ph-RE（546个氨基酸残基）编码的荧光素酶cDNA同源性为71%。Pv（GR）和Ph-RE酶与萤火虫的同源性分别为50 - 55%和46 - 49%，与金龟子的同源性为47 - 49%。Ph-RE荧光素酶具有一些独特的残基，取代了其他甲虫荧光素酶中的不变残基。在Pv（GR）多肽中缺失的Ph-RE中的额外残基Arg 352似乎是与红光产生相关的另一个重要结构特征。在研究的其他铁路蠕虫和点击甲虫荧光素酶的情况下，Phrixothrix荧光素酶不经历萤火虫荧光素酶在降低pH值时所遭受的典型红移，这可能与铁路蠕虫和点击甲虫荧光素酶之间共享的许多氨基酸残基有关。
Phrixothrix railroad-worms emit yellow-green light through 11 pairs of lateral lanterns along the body and red light through two cephalic lanterns. The cDNAs for the lateral lanterns luciferase of Phrixothrix vivianii, which emit green light (lambda(max) = 542 nm), and for the head lanterns of P. hirtus, which emit the most red-shifted bioluminescence (lambda(max) = 628 nm) among luminescent beetles, were cloned. Positive clones which emitted green (Pv(GR): lambda(max) = 549 nm) and red(Ph-RE: lambda(max) = 622 nm) bioluminescence were isolated. The lucifereases coded by Pv(GR) (545 amino acid residues) and Ph-RE (546 amino acid residues) cDNAs share 71% identity. Pv(GR) and Ph-RE luciferases showed 50-55% and 46-49% identity with firefly luciferases, respectively, and 47-49% with click-beetle luciferases. Ph-RE luciferase has some unique residues which replace invariant residues in other beetle luciferases. The additional residue Arg 352 in Ph-RE, which is deleted in Pv(GR) polypeptide, seems to be another important structural feature associated with red light production. As in the case of other railroad-worms and click-beetle luciferases studied, Phrixothrix luciferases do not undergo the typical red shift suffered by firefly luciferases upon decreasing pH, a property which might be related to the many amino acid residues shared in common between railroad-worm and click-beetle luciferase.