Differential effects of short affinity tags on the crystallization of Pyrococcus furiosus maltodextrin-binding protein.
Pyrococcus furiosus maltodextrin-binding protein readily forms large orthorhombic crystals that diffract to high resolution. This protein was used as a model system to investigate the influence of five short affinity tags (His(6), Arg(5), Strep tag II, FLAG tag and the biotin acceptor peptide) on the formation of protein crystals and their ability to diffract X-rays. The results indicate that the amino-acid sequence of the tag can have a profound effect on both of these parameters. Consequently, the ability to obtain diffracting crystals of a particular protein may depend as much on which affinity tag is selected as it does on whether an affinity tag is used at all.
Bucher
Matthew H
MH
Protein Engineering Section, Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute at Frederick, PO Box B, Frederick, MD 21702, USA.
Evdokimov
Artem G
AG
Waugh
David S
DS
D001426:Bacterial Proteins / Q000737:chemistry*; D002352:Carrier Proteins / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D029968:Escherichia coli Proteins; D008958:Models, Molecular; D033902:Periplasmic Binding Proteins; D011487:Protein Conformation; D020027:Pyrococcus furiosus / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry
D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; C461619:MalE protein, E coli; D033902:Periplasmic Binding Proteins; D011994:Recombinant Proteins