Methods for the analysis of HIV-1 nucleocapsid protein interactions with oligonucleotides.
HIV-1 Nucleocapsid protein (NC) is a small basic protein that contains two retroviral zinc fingers. It is a highly effective nucleic acid chaperone that plays a critical role in viral replication acting as a cofactor in reverse transcription as well as other aspects of the viral lifecycle. We have used a variety of biophysical techniques to characterize the high affinity binding of NC to a short deoxyoligonucleotide (d(TG)(4)). Here we outline in detail the use of fluorescence anisotropy and surface plasmon resonance spectroscopy to study the binding of NC to d(TG)(4).
Stephen
Andrew G
AG
Protein Chemistry Laboratory, Research Technology Program, SAIC-Frederick, Inc., NCI-Frederick, Frederick MD, USA.
Fisher
Robert J
RJ
D005454:Fluorescence Polarization / Q000379:methods; D015497:HIV-1 / Q000502:physiology*; D009841:Oligonucleotides / Q000378:metabolism*; D011485:Protein Binding; D020349:Surface Plasmon Resonance / Q000379:methods; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000378:metabolism*
C071884:NCP7 protein, Human immunodeficiency virus 1; D009841:Oligonucleotides; D054301:gag Gene Products, Human Immunodeficiency Virus