Purification of a factor that hydrolyzes [2-14C]-2,5'-anhydro-3'-azido-3'-deoxythymidine from human liver (Meeting abstract).

Important note: Information in this article was accurate in 1995. The state of the art may have changed since the publication date.

Purification of a factor that hydrolyzes [2-14C]-2,5'-anhydro-3'-azido-3'-deoxythymidine from human liver (Meeting abstract).

Proc Annu Meet Am Assoc Cancer Res; 35:A3245 1994. Unique Identifier : AIDSLINE ICDB/95604930
Sun AS; Prusoff WH; Dept. of Pharmacology, Yale University School of Medicine, New; Haven, CT 06510

Abstract: 2,5'-anhydro-3'-azido-3'-deoxythymidine (anAZT) has antiviral activity against HIV-1 in cell culture and is converted to AZT and then thymine (thy) by human liver homogenate in 0.1% Triton X-100. The reaction products were identified with SAX HPLC. Ninety percent of the activity was recovered in the 100,000 xg supernatant (S) with a 2.2-fold purification. Ethanol was added to S (9:1) and the supernatant (90S) was evaporated to 0.5 ml and what adhered to the glass was solubilized in 0.1% Triton X-100 and lyophilized to 1 ml. Recovery in 90S was 78% with 150-fold purification. 90S was further fractionated with Sephacryl S-300 column (150 x 2.5 cm) and eluted with 0.2% Triton X-100 and 0.15 N NaCl at 4 C. Total recovery was 75% with the peak fraction having 414-fold of purification with 8.6% recovery. Two other fractions containing 22% of original activity were lyophilized to 2 ml and further purified with isoelectric focusing from pH 3 to pH 9. Fractions after IEF were eluted with 0.1% Triton X-100, dialyzed with distilled water, and lyophilized to 0.5 ml. The major activity was recovered in the fraction at pH 4.3 with a recovery of 5.3%. The reaction seems to be catalyzed by a macromolecule(s) with a molecular weight greater than 100,000.

Keywords: Carbon Radioisotopes Chromatography, Gel Chromatography, High Pressure Liquid Human Hydrolysis Isoelectric Focusing Liver/*METABOLISM Thymine Nucleotides/*METABOLISM ABSTRACT
950330
M9530831

AEGiS is a 501(c)3, not-for-profit, tax-exempt, educational corporation. AEGiS is made possible through unrestricted funding from Boehringer Ingelheim, Bridgestone/Firestone Charitable Trust, Bristol-Myers Squibb Company, Elton John AIDS Foundation, Gill Foundation, the National Library of Medicine, Quest Diagnostics, Roche and Trimeris, and donations from users like you. Always watch for outdated information. This article first appeared in 1995. This material is designed to support, not replace, the relationship that exists between you and your doctor.

AEGiS presents published material, reprinted with permission and neither endorses nor opposes any material. All information contained on this website, including information relating to health conditions, products, and treatments, is for informational purposes only. It is often presented in summary or aggregate form. It is not meant to be a substitute for the advice provided by your own physician or other medical professionals. Always discuss treatment options with a doctor who specializes in treating HIV.

Copyright ©1980, 1995. AEGiS. All materials appearing on AEGiS are protected by copyright as a collective work or compilation under U.S. copyright and other laws and are the property of AEGiS, or the party credited as the provider of the content. .