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Title: A suite of engineered GFP molecules for oligomeric scaffolding

Applications ranging from synthetic biology to protein crystallization could be advanced by facile systems for connecting multiple proteins together in predefined spatial relationships. One approach to this goal is to engineer many distinct assembly forms of a single carrier protein or scaffold, to which other proteins of interest can then be readily attached. In this work we chose GFP as a scaffold and engineered many alternative oligomeric forms, driven by either specific disulfide bond formation or metal ion addition. We generated a wide range of spatial arrangements of GFP subunits from 11 different oligomeric variants, and determined their X-ray structures in a total of 33 distinct crystal forms. Furthermore, some of the oligomeric GFP variants show geometric polymorphism depending on conditions, while others show considerable geometric rigidity. Potential future applications of this system are discussed.
 [1] ;  [1] ;  [1] ;  [2] ;  [2] ;  [2] ;  [1]
  1. Univ. of California, Los Angeles, CA (United States)
  2. Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
Publication Date:
OSTI Identifier:
Report Number(s):
Journal ID: ISSN 0969-2126; PII: S0969212615002890
Grant/Contract Number:
P01 GM098177; T32GM007185; FC02-02ER63421; RR-15301; AC02-06CH11357; AC52-06NA25396
Accepted Manuscript
Journal Name:
Additional Journal Information:
Journal Volume: 23; Journal Issue: 9; Journal ID: ISSN 0969-2126
Research Org:
Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)
Sponsoring Org:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23); USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Country of Publication:
United States
59 BASIC BIOLOGICAL SCIENCES protein scaffolds; protein crystallization; protein engineering; protein assembly; synthetic biology