>> HOMEPAGE

AIDS Research

Effects of trace metal compounds on HIV-1 reverse transcriptase:
an in vitro study

Biol Trace Elem Res 1999 May;68(2):107-19 (ISSN: 0163-4984)

Rivas CI; Vera JC; Guaiquil VH; Velasquez FV; Borquez-Ojeda OA; Carcamo JG; Concha II; Golde DW Program in Molecular Pharmacology and Therapeutics, Memorial Sloan-Kettering Cancer Center, New York, New York 10021, USA.

The effect of 44 different metal ions (Ag+, Al3+, As(O-)2, Au3+, Ba2+, Be2+, Bi3+, Cd2+, Ce3+, CO2+, Cr(O2-)4, Cr3+, Cs+, Cu2+, Fe3+, Fe2+, Ga3+, Ge4+, Hg2+, Ir4+, La3+, Li+, Mn2+, MO6+, Ni2+, OS4+, Pb2+, Pt4+, Rb+, Rh3+, Sb5+, Se(O2-)4, Se(O2-)3, Sn2+, Sr2+, Th4+, T1+, U(O2+)2, V(O-)3, VO2+, W(O2-)4, Y3+, Zn2+, and Zr4+) on the activity of the reverse transcriptase (RT) of the human immunodeficiency virus (HIV-1) was investigated in vitro. For this study, the RT activity assay was carried out by means of an enzyme-linked immunosorbent assay (ELISA) kit, using the template/primer hybrid poly(A) oligo(dT)15, which required some modifications: (1) possible interfering metal chelators (such as EDTA) in the original lysis buffer were avoided, and a new buffer (50 mM Tris-NO3, pH 7.8) was used throughout; (2) an amount of 2 ng of RT per well was considered to be optimal after checking the linearity of the reaction with increasing amounts of enzyme; (3) an incubation temperature of 37 degrees C and an incubation time of 1 h were chosen after preliminary studies in a wide range of temperature and time. At an incubation temperature > or = 40 degrees C, there was a dramatic loss of enzymatic activity. In addition, when RT alone was preincubated for 1 h at 5 degrees C, 25 degrees C, and 37 degrees C, there was a large (83%) loss of activity at 37 C as compared to that at 5 degrees C. These results are indicative of enzyme thermolability, which is higher in the absence of substrates. The effect of metal ions on RT activity was tested using two different metal salt concentrations (10(-4) M and 10(-5) M). Under such experimental conditions, the presence of five metal ions (Pt4+, Ag+, Rh3+, Zn2+, and Hg2+) decreased the RT activity in a dose-response fashion. The observed order of effectiveness with respect to inhibition was Pt4+ > Ag+ > Rh3+ > Zn2+ = Hg2+. Estimated mean inhibitory concentrations (IC50) were 7.8 microM for (NH4)2PtCl6, 14.1 microM for AgNO3, 46.8 microM for RhCl3, 53.7 microM for Zn(SO)4, and 56.2 microM for Hg(NO3)2. Because these data are of the same order of magnitude as the corresponding values related to other RT inhibitors used in anti-AIDS therapy, metal compounds or their derivatives could give an interesting contribution in the development of new RT inhibitors for clinical use.

Sitemap

Was ist neu:

Ignaz Semmelweis: Zuerst von seinen Kollegen belächelt, später als Held gefeiert

Beenden Sie die Verdammung der Naturheilkunde! – Offener Brief von Dr. Matthias Rath an Bundeskanzlerin Angela Merkel

Prof. Edzard Ernst: Karriere durch Verunglimpfung von Naturheilverfahren?

Für eine gerechtere Welt – ohne Armut und Volkskrankheiten

William Harvey – ein Arzt der verstand, dass er seiner Zeit voraus war

Herber Rückschlag für die Biotech-Industrie: Insekten entwickeln Resistenzen gegenüber Gen-Pflanzen

» Natürliche Gesundheit

» "Geschäft m. d. Krankheit"

» Internationale Kampagne

» Dr. Raths Offene Briefe

» Studien weltweit

» Studien auf Deutsch

» Helfen Sie mit!

» Codex Alimentarius

» Dossiers

» Sitemap

Dr. Rath Health Foundation auf Facebook Dr. Rath Health Foundation auf YouTube Dr. Rath Health Foundation auf Twitter RSS
 © 2017 Dr. Rath Health Foundation | Kontakt | Impressum | RSS Feed