Protein purification with C-terminal fusion of maltose binding protein.
For affinity-chromatography-based purification of proteins that are prone to abnormal termination of translation or that may not be modified at their N-termini, affinity tags are needed which can be fused to the C-terminus. In this publication we describe that maltose binding protein (MBP) fused to the C-terminus of the plant photoreceptor phytochrome B allows purification of the fusion protein via amylose affinity chromatography. After overexpression in yeast a 125-fold enrichment could be achieved. The spectral properties of phytochrome B were not impaired by the fusion and purification. These results demonstrate that not only the widely used N-terminal fusions of MBP but also C-terminal fusions can be employed for protein purification.
Hennig
L
L
Institut für Biologie II, Universität Freiburg, Schänzlestrasse 1, Freiburg, 79104, Germany.
Schäfer
E
E
D000345:Affinity Labels; D000688:Amylose; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics; D002846:Chromatography, Affinity / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D008320:Maltose; D058619:Maltose-Binding Proteins; D010786:Photoreceptor Cells; D010834:Phytochrome / Q000302:isolation & purification*; D051939:Phytochrome B; D010947:Plants, Toxic; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013053:Spectrophotometry; D014026:Nicotiana / Q000235:genetics; D014157:Transcription Factors
D000345:Affinity Labels; D001426:Bacterial Proteins; D002352:Carrier Proteins; D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D010834:Phytochrome; D051939:Phytochrome B; D008320:Maltose; D000688:Amylose