Measure of O(6)-Alkylguanine-DNA Alkyltransferase Activity in Normal Human Epidermal Keratinocytes in Culture and Effects of Bis-(2-chloroethyl)sulfide in the Activity.
Abstract
O(sup 6)-Alkylguanine-DNA alkyltransferase (AGT) is a DNA repair protein that removes alkyl adducts from O(sup 6)-alkylguanine in DNA. AGT may be important in DNA repair following injury induced by bis-(2-chloroethyl)sulfide (sulfur mustard, HD), since O(sup 6)-alkylguanine is one of the HD alkylation products. One model for HD-induced injury uses normal human epidermal keratinocytes (NHEK) in culture. In this study we measured the levels of AGT in NHEK grown to either 60 - 80% or to 100% confluency and then studied the effects of a one-hour exposure to 50, 100, and 300 micro-M HD on AGT activity. Mean AGT activity in NHEK grown to 60.89% and 100% confluency was 470 +/- 404 and 518 +/- 737 fmol/mg protein, respectively. In general, AGT activity appeared to increase after exposure to 100 micro-M HD and decrease with increased confluency and after exposure to 50 and 300 micro-M HD. However, a two-way analysis of variance for cell confluency and HD concentration showed no significant differences between cell confluencies or the different HD concentrations.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 01, 1999
- Accession Number
- ADA366607
Entities
People
- Agenes Gaweska
- Charise C. Davenport
- Ellen M. Kurt
- Vincent M. O'donnell
Organizations
- United States Army Medical Research Institute of Chemical Defense