Open AccessPhotoaffinity Labeling of Chromatin
Author(s) -
NIELSEN Peter Eigil
Publication year - 1982
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1982.tb05878.x
Subject(s) - chromatin , photoaffinity labeling , histone , chemistry , dna , reagent , macromolecule , biophysics , biochemistry , stereochemistry , binding site , biology , organic chemistry
The syntheses and properties of potential photoaffinity labels for the proteins in chromatin studies are described. One compound, N , N '‐bis‐(9‐acridinyl)‐4‐aza‐4‐(4‐azidobenzoyl)‐1,7‐diaminoheptane dihydrochloride, has been studied more closely. This photolabel shows high affinity towards DNA (K a ∼ 3 × 10 5 M −1 ) and photoreacts with histones at wavelengths in the range 260–450 nm. The photoreaction was monitored fluori‐ metrically, and labeling of histones H1, H2A/H2B and H3 was observed. When chromatin was photolabeled, labeling of H1 was exclusively quenched by NaCl at concentrations which are known to cause dissociation of this histone from the DNA. It is thus inferred that only DNA‐associated proteins are photolabeled by the reagent.