Lund S G, Dyment P, Gervais M R, Moyes C D, Tufts B L
Department of Biology, Queen's University, Kingston, Ontario, Canada K7L 3N6.
J Comp Physiol B. 2002 Aug;172(6):467-76. doi: 10.1007/s00360-002-0269-9. Epub 2002 Jun 29.
This study investigates the evolutionary history of vertebrate red blood cell carbonic anhydrase (CA) by characterizing the isozyme properties and nucleotide sequence of an ancient fish, the longnose gar ( Lepisosteus osseus). The inhibitor sensitivities of gar rbc CA closely resembled those for mammalian CA II, as well as those for CAs from more recently evolved fishes. The kinetic properties of gar rbc CA were not closely aligned with either mammalian CA I and CA II, but fit well into an emerging phylogenetic pattern for early vertebrates. Gar rbc CA cDNA was also amplified from mRNA using 5' and 3'-RACE and the open reading frame consisted of 786 bp. This sequence shares approximately 65% identity with the nucleotide and amino acid sequences of both mammalian CA I and CA II. When the amino acid sequences within the active site are compared, gar rbc CA differs from mammalian CA I, CA II and CA VII by 9, 4 and 3 of the 36 amino acids, respectively. Phylogenetic analyses suggest that gar rbc CA diverged before the amniotic CAs (CA I, CA II and CA III), but after CA V and CA VII.
本研究通过对一种古老鱼类——长吻雀鳝(Lepisosteus osseus)的同工酶特性和核苷酸序列进行表征,来探究脊椎动物红细胞碳酸酐酶(CA)的进化历史。雀鳝红细胞CA的抑制剂敏感性与哺乳动物CA II以及较近期进化的鱼类的CA的抑制剂敏感性非常相似。雀鳝红细胞CA的动力学特性与哺乳动物CA I和CA II均不紧密匹配,但很好地符合早期脊椎动物新出现的系统发育模式。还利用5'和3'-RACE从mRNA中扩增出雀鳝红细胞CA cDNA,其开放阅读框由786 bp组成。该序列与哺乳动物CA I和CA II的核苷酸序列及氨基酸序列均具有约65%的同一性。当比较活性位点内的氨基酸序列时,雀鳝红细胞CA与哺乳动物CA I、CA II和CA VII在36个氨基酸中分别有9个、4个和3个氨基酸不同。系统发育分析表明,雀鳝红细胞CA在羊膜动物的CA(CA I、CA II和CA III)之前分化,但在CA V和CA VII之后。