Book chapter
Expression, Purification, and Biochemical Evaluation of Human RAD51 Protein
Methods in enzymology, pp.157-178
Methods in Enzymology, Elsevier
01/01/2018
DOI: 10.1016/bs.mie.2017.11.011
PMCID: PMC6033273
PMID: 29458757
Abstract
The RAD51 DNA strand exchange protein plays an important role in maintaining the integrity of the human genome. It promotes homology-directed DNA repair by exchanging strands between the damaged and the intact DNA molecules. It also plays an important role in stabilizing distressed DNA replication forks. When overexpressed or mis-regulated, however, RAD51 contributes to "rogue," genome destabilizing events that can lead to cancer, cell death, and to acquisition of chemotherapy resistance by cancerous cells. Human RAD51 is, therefore, an important and highly coveted anticancer drug target. Biochemical, biophysical, and structural studies of the human RAD51 and establishment of its structure-activity relationship require purification of large quantities of protein. In this chapter we describe a robust method for expression and purification of human RAD51 and the methods for assessing its activity based on the single-strand DNA-binding stoichiometry and its capacity to carry out the DNA strand exchange reaction.
Details
- Title: Subtitle
- Expression, Purification, and Biochemical Evaluation of Human RAD51 Protein
- Creators
- Shyamal Subramanyam - Roy J. and Lucille A. Carver College of MedicineMaria Spies - Roy J. and Lucille A. Carver College of Medicine
- Contributors
- A Malkova (Editor)M Spies (Editor) - University of Iowa, Biochemistry and Molecular Biology
- Resource Type
- Book chapter
- Publication Details
- Methods in enzymology, pp.157-178
- Publisher
- Elsevier; SAN DIEGO
- Series
- Methods in Enzymology
- DOI
- 10.1016/bs.mie.2017.11.011
- PMID
- 29458757
- PMCID
- PMC6033273
- eISSN
- 1557-7988
- ISSN
- 0076-6879
- Number of pages
- 22
- Grant note
- P30 CA086862 / NCI NIH HHS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Cancer Institute (NCI) P30CA086862 / NATIONAL CANCER INSTITUTE; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Cancer Institute (NCI) R01 GM108617 / NIGMS NIH HHS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS) R01GM108617 / NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS)
- Language
- English
- Date published
- 01/01/2018
- Academic Unit
- Radiation Oncology; Biochemistry and Molecular Biology
- Record Identifier
- 9984293090002771
Metrics
17 Record Views