XTT formazan widely used to detect cell viability inhibits HIV type 1 infection in vitro by targeting gp41.
XTT can be metabolically reduced by mitochondrial dehydrogenase in viable cells to a water-soluble formazan product. Thus XTT has been widely used to evaluate cell viability and to screen anti-HIV agents and the cytotoxicity of these agents. The present studies demonstrated that XTT formazan derived from XTT in cell culture significantly inhibits the fusion of HIV-1-infected cells with uninfected cells. Synthetic XTT formazan effectively inhibited the replication of laboratory-adapted and primary HIV-1 isolates and cell-to-cell fusion with low cytotoxicity. It blocks the six-helix bundle formation between peptides derived from the N- and C-terminal heptad repeat regions of the gp41 ectodomain (designated N- and C-peptides, respectively). Analysis by a computer-aided docking program indicates that XTT formazan may bind to the highly conserved hydrophobic pocket on the surface of the central trimeric coiled coil of gp41. These results suggest that XTT formazan inhibits HIV-1 entry by targeting the alpha-helical coiled-coil domain of gp41. This small molecular nonpeptide antiviral compound can be used as a lead for designing more effective HIV-1 entry inhibitors targeting the fusion stage of HIV-1 infection. But because XTT formazan itself has anti-HIV-1 activity, caution should be exercised when XTT is used to evaluate HIV-1 infectivity.
Zhao
Qian
Q
Lindsley F. Kimball Research Institute, New York Blood Center, New York, New York 10021, USA.
Ernst
Justin T
JT
Hamilton
Andrew D
AD
Debnath
Asim K
AK
Jiang
Shibo
S
D000818:Animals; D001665:Binding Sites; D002459:Cell Fusion; D002460:Cell Line; D002470:Cell Survival; D002942:Circular Dichroism; D005562:Formazans / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D015700:HIV Envelope Protein gp41 / Q000737:chemistry / Q000378:metabolism; D015658:HIV Infections / Q000821:virology*; D015497:HIV-1 / Q000187:drug effects* / Q000472:pathogenicity*; D006801:Humans; D008561:Membrane Fusion / Q000187:drug effects; D008773:Methylphenazonium Methosulfate / Q000494:pharmacology; D051379:Mice; D008958:Models, Molecular; D014779:Virus Replication / Q000187:drug effects*
D005562:Formazans; D015700:HIV Envelope Protein gp41; D008773:Methylphenazonium Methosulfate; C059088:1,5-bis(2-methoxy-4-nitro-5-sulfophenyl)-3-((phenylamino)carbonyl)formazan