Thiol protection in membrane protein purifications: a study with phage holins. Academic Article

abstract

• The lambda holin, or S105, is a small cytoplasmic membrane protein that controls the timing of host lysis. Using thiol-specific reagents, we determined that the single cysteine residue within S105 was heterogeneously modified during membrane extraction and subsequent immobilized metal ion chromatography. Here we describe the use of a specific and reversible thiol reagent, 2,2'-dithiodipyridine, to generate purified protein with its cysteine residues in the native thiol state. The 2,2'-dithiodipyridine protection protocol was also successfully used for another unrelated holin, S(21)68, and should be generally useful for the purification of membrane proteins.

authors

• Young, Ryland

published proceedings

• Anal Biochem

author list (cited authors)

• Dewey, J. S., Struck, D. K., & Young, R. y.

citation count

• 3

complete list of authors

• Dewey, Jill S||Struck, Douglas K||Young, Ry

publication date

• January 2009

publisher

• Elsevier  Publisher

published in

• Analytical Biochemistry  Journal

keywords

• 2,2'-Dipyridyl
• Amino Acid Sequence
• Bacteriophages
• Cysteine
• Disulfides
• Membrane Proteins
• Molecular Sequence Data
• Viral Proteins

Digital Object Identifier (DOI)

• 10.1016/j.ab.2009.04.031

start page

• 221

end page

• 223

volume

• 390

issue

• 2

URL

• http://dx.doi.org/10.1016/j.ab.2009.04.031