Abstract
Abstract: :
Purpose: To functionaly compare retina specific amine oxidase (AOC2) with super family gene vascular adhesion protein 1 (AOC3). Methods: The mouse retina specific amine oxidase (AOC2) previously cloned by cDNA library screening and 5'/3'-RACE method was used as probe to screen mouse genomic library. The entire genomic structure was deterimed and the 5' and 3' regions beyond the AOC2 gene were also sequenced. AOC2 promoter sequence was analyzed with trascription factor database and compared with AOC3 promoter. The coding sequence of mouse AOC2 cDNA was cloned into expression vectors (pMIB/V5-His, Invitrogen, Inc.) for Sf21 insect cell expression. Purified protein was tested for diamine oxidase acitivty using putrescine as substrate. The 3D structure of AOC2 protein was created by 3D modeling software based on bacteria amine oxidase crystal structure and compared with AOC3. Results: The AOC2, 4,625 bp in legnth, is composed of 4 exons and 3 introns following the canonical donor-acceptor rule. One of the super family of amine oxidase, vascular adhesion protein 1 (AOC3), was tandemly located approximately 1.4 Kb downstream of AOC2 exon 4. This was also confirmed in human gene structure of AOC2 and AOC3. Promoter sequence comparison of AOC2 and AOC3 revealed significantly low similarity (10%) but several short homologous stretch of sequences matching but in different order. Comparison of AOC2 and AOC3 protein structure showed posible functional similarity. Conclusion: Super family of aime oxidase AOC2 and AOC3 was tandemly located approximately 1.4 Kb apart. Enzyme activity and 3D structure analysis show functional similarity but different tissue specificity.
Keywords: 415 ganglion cells • 557 retina: proximal(bipolar, amacrine, and ganglion cells) • 476 molecular biology