Analysis of Keratin Filament Assembly/Disassembly and Structure Following Modification by Sulfur Mustard Analogs
Abstract
Characterization of vimentin intermediate filament (IF) structure by site directed spin labeling (SDSL) and electron paramagnetic resonance (EPR) has produced new data concerning the All arrangement of molecules in intact filaments. In the All alignement, rod 1B of a dimer is aligned with rod 1B of a second dimer. Our EPR data identify vimentin position 190 as the midpoint of overlap between dimers in the A11 arrangement. In conjunction with previously published data, we have thus identified contact surfaces between individual amino acids of rod 1B and rod 2B, involved in assembly of IFs. Analysis of keratin filament assembly and disassembly following treatment with the vesicant analogs 2-chloroethyl ethyl sulfide (CEES) and mechlorethamine (MEC) has been performed with 2 keratin pairs, one bacterially produced and one isolated from bovine tissue. Experiments with bacterially produced keratin 8 and 18 show that Intermediate Filaments assembled in vitro and subsequently treated with CEES and MEC are severely altered. The normal filament network is destroyed, replaced by aggregates of material with an irregular appearance. Experiments with k5 and k14 isolated from bovine tissue reveal the same results; CEES and MEC treatment destroy the smooth filamentous appearance of normal IFs and result in large aggregates.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 2003
- Accession Number
- ADA418634
Entities
People
- John F. Hess
Organizations
- University of California