Targeting cysteine residues of human immunodeficiency virus type 1 protease by reactive free radical species.
Nitric oxide (NO) is a naturally occurring free radical with many functions. The oxidized form of NO, the nitrosonium ion, reacts with the thiol group of cysteine residues resulting in their modification to S-nitrosothiols. The human immunodeficiency virus type 1 (HIV-1) protease (HIV-PR) has two cysteine residues that are conserved amongst different viral isolates found in patients with acquired immunodeficiency syndrome (AIDS). In an active dimer, these residues are located near the surface of the protease. We have found that treatment of HIV-PR with different NO congeners results in loss of its proteolytic activity and simultaneous formation of S-nitrosothiols. Sodium nitroprusside inhibited HIV-PR up to 70% and S-nitroso-N-acetylpenicillamine completely inhibited the protease within 5 min of treatment. The pattern of inhibition by NO donors is comparable to its inhibition by N-acetyl pepstatin. Using electrospray ionization-mass spectrometry, we identified the modification of HIV-PR by NO as that of S-nitrosation. Our findings point towards a possible role of NO in mediating resistance to HIV-1 infection.
Department of Biochemistry, Weill Medical College of Cornell University, New York, New York 10021, USA.
A
A
Basu
P K
PK
Sehajpal
J S
JS
Ogiste
H M
HM
Lander
D001419:Bacteria / Q000201:enzymology / Q000235:genetics; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D016333:HIV Protease / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D017320:HIV Protease Inhibitors / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D002479:Inclusion Bodies / Q000201:enzymology; D009569:Nitric Oxide / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D009599:Nitroprusside / Q000494:pharmacology; D010396:Penicillamine / Q000031:analogs & derivatives / Q000494:pharmacology; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D026423:S-Nitroso-N-Acetylpenicillamine; D021241:Spectrometry, Mass, Electrospray Ionization; D014665:Vasodilator Agents / Q000494:pharmacology
D017320:HIV Protease Inhibitors; D020030:Nitric Oxide Donors; D011994:Recombinant Proteins; D014665:Vasodilator Agents; D009599:Nitroprusside; D009569:Nitric Oxide; D026423:S-Nitroso-N-Acetylpenicillamine; D016333:HIV Protease; D010396:Penicillamine; D003545:Cysteine