Linear dichroism of the complex between the gene 32 protein of bacteriophage T4 and poly(1,N6-ethenoadenylic acid).
We performed linear dichroism measurements in compressed polyacrylamide gels on the complex between the helix-destabilizing protein of bacteriophage T4, GP32 and poly(1,N6-ethenoadenylic acid), which is used as a model system for single-stranded DNA. A strong hyperchromism for poly(1,N6-ethenoadenylic acid) in the complex indicates a strongly altered conformation. The positive linear dichroism in the wavelength region where the bases absorb must be explained by a strong tilting of the bases in the complex. This finding is in accordance with results from earlier studies, using electric birefringence and circular dichroism measurements. Our measurements show that the angle between the bases and the local helix axis is 42(+/- 6)degrees. In addition, a pronounced contribution from the tryptophan residues of GP32 can be recognized, indicating that several of these residues have a specific orientation in the complex. The sign of the dichroism due to the tryptophan residues is the same as that due to the DNA bases. However, it is not sufficient to assume that all the observed dichroism is due to one or more intercalated tryptophan residues and there must be one or more additional tryptophan residues that make an angle of less than 40 degrees with the local helix axis. Some possible structures of the DNA-protein complex are discussed.
van Amerongen
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H
Department of Biophysics, Physics Laboratory of the Free University, Amsterdam, The Netherlands.
Kuil
M E
ME
van Mourik
F
F
van Grondelle
R
R
D004268:DNA-Binding Proteins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005814:Genes, Viral; D046911:Macromolecular Substances; D011061:Poly A / Q000378:metabolism*; D013057:Spectrum Analysis; D013604:T-Phages / Q000235:genetics*; D014364:Tryptophan; D014764:Viral Proteins / Q000378:metabolism*
D004268:DNA-Binding Proteins; D046911:Macromolecular Substances; D014764:Viral Proteins; D011061:Poly A; C005163:poly(1,N(6)-ethenoadenylic acid); D014364:Tryptophan