4.2 Article

Expression, purification, and characterization of proteins from high-quality combinatorial libraries of the mammalian calmodulin central linker

期刊

PROTEIN EXPRESSION AND PURIFICATION
卷 75, 期 2, 页码 186-191

出版社

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.pep.2010.08.007

关键词

Protein design; Combinatorial library; Binary patterning; Protein engineering

资金

  1. University of Kentucky Office of Undergraduate Research and Creative Activities/eUreKa
  2. NIH National Center for Research Resources (NCRR) [P20 RR020171]
  3. Kentucky Science and Engineering Foundation with the Kentucky Science and Technology Corporation [KSEF-148-502-07-201]
  4. University of Kentucky College of Medicine

向作者/读者索取更多资源

Combinatorial libraries offer an attractive approach towards exploring protein sequence structure and function Although several strategies introduce sequence diversity the likelihood of identifying proteins with novel functions is increased when the library of genes encodes for folded and soluble structures Here we present the first application of the binary patterning approach of combinatorial protein library design to the unique central linker region of the highly-conserved protein calmodulin (CaM) We show that this high quality approach translates very well to the CaM protein scaffold All library members over-express and are functionally diverse having a range of conformations in the presence and absence of calcium as determined by circular dichroism spectroscopy Collectively these data support that the binary patterning approach when applied to the highly-conserved protein fold can yield large collections of folded soluble and highly-expressible proteins (C) 2010 Elsevier Inc All rights reserved

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