Biotinylation of proteins in solution and on cell surfaces

Current Protocols in Protein Science
E J Luna

Abstract

This unit contains three protocols for biotinylation of isolated proteins: attaching biotin to primary amines (e.g., amino groups of lysyl residues); attaching biotin to sulfhydryls (i.e., thiol groups of cysteinyl residues); and attaching biotin to carbohydrate residues on proteins. As biotinylation of lysyl and cysteinyl residues may alter protein function, modification of protein carbohydrates, which is usually innocuous, may be preferable if intact protein function is required (e.g., for activity assays or affinity purification). Biotinylation of cysteinyl thiols requires that disulfide bonds in isolated proteins be reduced before labeling. Biotinylation of surface proteins on living cells is also described using mild reaction conditions. Finally, this unit includes a brief description of methods for detecting biotinylated proteins.

References

Jan 1, 1975·Advances in Protein Chemistry·N M Green
Apr 15, 1979·Analytical Biochemistry·K P HenriksonW L Maloy
Jan 1, 1990·Methods in Enzymology·J P Kraehenbuhl, C Bonnard
Jun 27, 1990·Journal of Chromatography·E A Bayer, M Wilchek
Jan 1, 1990·Methods in Enzymology·R A Kohanski, M D Lane
Aug 16, 1990·Biochemical and Biophysical Research Communications·R AlonM Wilchek
Jul 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·H M IngallsE J Luna
Jan 1, 1987·Methods in Cell Biology·C M Goodloe-Holland, E J Luna
Jan 1, 1985·Annals of the New York Academy of Sciences·R A Kohanski, M D Lane
Apr 1, 1993·Molecular Biology Reports·G MattsonS Morgensen

❮ Previous
Next ❯

Citations

Feb 12, 2008·Current Protocols in Molecular Biology·D G Edmondson, S Y Roth
Jan 30, 2008·Current Protocols in Cell Biology·D G Edmondson, S Y Roth

❮ 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.