Labelling of histone H5 and its interaction with DNA. 1. Histone H5 labelling with fluorescein isothiocyanate

Journal of Biomolecular Structure & Dynamics
M FavazzaC Houssier

Abstract

Histone H5 has been labelled with fluorescein isothiocyanate (FITC) with particular attention to the reaction conditions (pH, reaction time and input FITC/H5 molar ratio) and to the complete elimination of non-covalently bound dye. We preferred to use reaction conditions which yielded non-specific uniform labelling rather than specific alpha-NH2 terminal labelling, in order to obtain higher sensitivity in further studies dealing with the detection of perturbation at the binding sites of H5 on DNA. FITC-labelled H5 was further characterized by absorption and circular dichroism spectroscopy, and the fluorescein probe titrated in the 4-8 pH range. The structural integrity of H5 was found to be preserved after labelling. The positive electrostatic potential of the environment in which the FITC probe is embedded in the arginine/lysine-rich tails of H5 is believed to be responsible for the drop of pK of 1 unit found for H5-FITC as compared to free FITC. For the globular part of H5, the pK of covalently-bound FITC was only slightly lowered; this is a consequence of the much lower content in positively-charged amino-acid side chains in this region.

References

Jul 1, 1978·Nucleic Acids Research·L WynsR Hamers
Jan 1, 1978·Cold Spring Harbor Symposia on Quantitative Biology·A E DieterichC R Cantor
Jan 1, 1978·Annual Review of Biochemistry·L Stryer
Apr 25, 1979·Journal of Molecular Biology·A E DieterichC R Cantor
Jun 19, 1979·Biochimica Et Biophysica Acta·F J AvilesE M Bradbury
Aug 1, 1978·European Journal of Biochemistry·F J AvilesE M Bradbury
Oct 4, 1977·Biochimica Et Biophysica Acta·K Javaherian, S Amini
Aug 16, 1976·European Journal of Biochemistry·C Crane-RobinsonM Daune
Apr 25, 1986·Nucleic Acids Research·F Watanabe
Dec 1, 1988·European Journal of Biochemistry·D J Clark, J O Thomas
Sep 1, 1987·European Journal of Biochemistry·V R RussanovaI G Pashev
Apr 22, 1986·Biochemistry·D G Chung, P N Lewis
Feb 20, 1986·Journal of Molecular Biology·D J Clark, J O Thomas
Oct 1, 1986·Cancer Genetics and Cytogenetics·J A ReyJ Benítez
Apr 1, 1986·European Journal of Biochemistry·L De PetrocellisG Geraci
Jul 27, 1966·Biochimica Et Biophysica Acta·A Jobbágy, K Király
May 1, 1967·Journal of Chromatography·J C Janson
Jan 1, 1983·Biopolymers·C Crane-Robinson, P L Privalov
May 7, 1984·FEBS Letters·F Watanabe
Jan 1, 1982·Annual Review of Biophysics and Bioengineering·L StryerC F Meares
Jan 1, 1981·Biopolymers·A E Dieterich, C R Cantor

❮ Previous
Next ❯

Citations

Jun 9, 2005·Pediatric Nephrology : Journal of the International Pediatric Nephrology Association·Gabriela C FernándezMarina S Palermo
Sep 30, 2000·European Journal of Biochemistry·J M HolopainenP K Kinnunen
Mar 30, 1999·European Journal of Biochemistry·J CaldenteyD H Bamford

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.