title: Nonhistone chromosomal protein HMG 1 interactions with DNA. Fluorescence and thermal denaturation studies.
abstract: The interaction of high mobility group protein 1 (HMG 1) isolated from chicken erythrocytes with DNA has been characterized using the intrinsic tryptophan fluorescence of the protein as a probe. It was found that the fluorescence is quenched approximately 30% upon binding to either single- or double-stranded DNA. Fluorescent titrations indicate that the physical site size for HMG 1 binding on native DNA is approximately 14 base pairs (or 14 bases for binding to single-stranded DNA). Binding to single-stranded poly(dA) is only slightly dependent on ionic strength, although the affinity for double-stranded DNA is strongly ionic strength-dependent and has an optimum at approximately 100-120 mM Na+. Above this range, binding to native DNA is virtually all electrostatic in nature. Although the affinity of HMG 1 for single-stranded DNA is higher than that for double-stranded DNA at the extremes of the ionic range studied, no clear evidence for a helix-destabilizing activity was obtained. At low ionic strength, the protein actually stabilized DNA against thermal denaturation, while at high ionic strength, HMG 1 appears to undergo denaturation below the Tm of the DNA. Studies of the environment of the tryptophan fluorophores using collisional quenchers iodide, cesium, and acrylamide suggest that the predominant fluorophore is relatively exposed but constrained in a rigid, positively charged environment.
mesh_terms: D000818:Animals; D001482:Base Composition; D002467:Cell Nucleus / Q000378:metabolism*; D002645:Chickens; D004247:DNA / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism; D006609:High Mobility Group Proteins / Q000097:blood* / Q000302:isolation & purification; D007700:Kinetics; D008970:Molecular Weight; D009691:Nucleic Acid Denaturation; D011485:Protein Binding; D011489:Protein Denaturation; D013050:Spectrometry, Fluorescence
chemical_list: D006609:High Mobility Group Proteins; D004247:DNA