A systematic approach to isolate mono-disperse membrane proteins - purification of zinc transporter ZntB.
Obtaining mono-disperse and stable protein is a requirement for successful structural and biochemical investigation of proteins. For membrane proteins, such preparation is one of the major hurdles, which consequently has contributed to the slow progress in studying them. During the past few years, many screening methods have been developed to make studies of membrane proteins more efficient. Despite these advances, many membrane proteins remain challenging to even isolate in a stable and homogeneous form. The bacterial zinc transporter ZntB is such a protein, for which no isolation procedure has been reported. Here, we present a systematic approach to obtain homogeneous and mono-disperse zinc transporter ZntB in quantities sufficient for structural and biochemical studies. Important aspects of this study that can be applied to other membrane proteins are also discussed.
Centre for Biomedical Structural Biology, School of Biological Sciences, Nanyang Technological University, Singapore.
Sally Thanh
ST
Van Pham
Henrik
H
Engman
Lars-Göran
LG
Dahlgren
Tobias
T
Cornvik
Said
S
Eshaghi
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D003001:Cloning, Molecular / Q000379:methods*; D003902:Detergents; D004926:Escherichia coli / Q000235:genetics*; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D012486:Salmonella typhimurium / Q000737:chemistry; D016415:Sequence Alignment; D012995:Solubility
D001426:Bacterial Proteins; D002352:Carrier Proteins; D003902:Detergents; D008565:Membrane Proteins; C017800:zinc-binding protein