Gene expression response to misfolded protein as a screen for soluble recombinant protein.
Proper protein folding is key to producing recombinant proteins for structure determination. We have examined the effect of misfolded recombinant protein on gene expression in Escherichia coli. Comparison of expression patterns indicates a unique set of genes responding to translational misfolding. The response is in part analogous to heat shock and suggests a translational component to the regulation. We have further utilized the expression information to generate reporters responsive to protein misfolding. These reporters were used to identify properly folded recombinant proteins and to create soluble domains of insoluble proteins for structural studies.
Lesley
Scott A
SA
Genomics Institute of the Novartis Research Foundation, 3115 Merryfield Row, San Diego, CA 92121, USA. lesley@gnf.org
Graziano
Jim
J
Cho
Charles Y
CY
Knuth
Mark W
MW
Klock
Heath E
HE
D000818:Animals; D015870:Gene Expression; D020869:Gene Expression Profiling; D017930:Genes, Reporter / Q000235:genetics; D006360:Heat-Shock Proteins / Q000378:metabolism*; D018869:Heat-Shock Response / Q000502:physiology*; D006358:Hot Temperature; D006801:Humans; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry* / Q000235:genetics*; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D012270:Ribosomes / Q000737:chemistry*; D012995:Solubility; D014158:Transcription, Genetic
D006360:Heat-Shock Proteins; D011506:Proteins; D011994:Recombinant Proteins