Molecular modeling and in vitro activity of an HIV-1-encoded glutathione peroxidase

摘要

Based on theoretical evidence. it has been proposed that HIV-1 may encode several selenoprotein modules. one of which (overlapping the env gp41-coding region) has highly significant sequence sim- ilarity to the mammalian selenoprotein glutathione peroxidase (GPx. EC 1.11.1.9). The similarity score of the putative HIV-1 viral GPx homolog relative to an aligned set of known GPx is 6.3 SD higher than expected for random sequences of similar composi- tion. Based an that alignment. a molecular model of the HIV-1 GPx was constructed by homology modeling from the bovine GPx crystal structure. Despite extensive truncation relative to the cellular GPx gene, the structural core and the geometry of the catalytic triad of selenocysteine, glutamine, and tryptophan are well conserved in the viral GPx. All of the insertions and deletions predicted by the alignment proved to be structurally feasible. The model is energetically favorable, with a computed molecular me- chanics strain energy close to that of the bovine GPx structure, when normalized on a per-residue basis. However, considering the remote homology, this model is intended only to provide a work- ing hypothesis allowing for a similar active site and structural core. To validate the theoretical predictions, we cloned the hypothetical HIV-1 gene and found it to encode functional GPx activity when expressed as a selenoprotein in mammalian cells. In transfected canine kidney cells, the increase in GPx activity ranged from 21/100 to 43/100 relative to controls (average 30/100, n = 9, P < 0.0001). whereas. in transfected MCF7 cells. which have low endogenous GPx activity, a near 100/100 increase was observed (average 99/100, n = 3, P < 0.05).