Abstract:
:Structural genomics has the ambitious goal of delivering three-dimensional structural information on a genome-wide scale. Yet only a small fraction of natural proteins are suitable for structure determination because of bottlenecks such as poor expression, aggregation, and misfolding of proteins, and difficulties in solubilization and crystallization. We propose to overcome these bottlenecks by producing soluble, highly expressed proteins that are derived from and closely related to their natural homologs. Here we demonstrate the utility of this approach by using a green fluorescent protein (GFP) folding reporter assay to evolve an enzymatically active, soluble variant of a hyperthermophilic protein that is normally insoluble when expressed in Escherichia coli, and determining its structure by X-ray crystallography. Analysis of the structure provides insight into the substrate specificity of the enzyme and the improved solubility of the variant.
journal_name
Nat Biotechnoljournal_title
Nature biotechnologyauthors
Pédelacq JD,Piltch E,Liong EC,Berendzen J,Kim CY,Rho BS,Park MS,Terwilliger TC,Waldo GSdoi
10.1038/nbt732subject
Has Abstractpub_date
2002-09-01 00:00:00pages
927-32issue
9eissn
1087-0156issn
1546-1696pii
nbt732journal_volume
20pub_type
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