Coiled coil miniprotein randomization on phage leads to charge pattern mimicry of the receptor recognition determinant of interleukin 5

Li, Chuanzhao; Plugariu, C.; Bajgier, Joanna; White, John R.; Liefer, Kristin M.; Wu, Sheng‐Jiun; Chaiken, Irwin M.

doi:10.1002/jmr.558

Cited by 6 publications

(2 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…9 -11 Each bundle is composed of three helices (A-C) from one chain and one (D) from the other. The structure reveals two ␤-strand regions each composed of residues 88 EERRR 92 between the C-and D-helices of each four-helix bundle and an antiparallel ␤-strand from the same bundle containing another charged residue Arg 32 . Site-directed mutagenesis, as well as other studies have shown the importance of charged residues in the CD turn (in particular Glu 89 and Arg 91 ) for receptor ␣ chain binding.…”

Section: Introductionmentioning

confidence: 99%

Cyclic peptide interleukin 5 antagonists mimic CD turn recognition epitope for receptor α

et al. 2004

Self Cite

View full text Add to dashboard Cite

The cyclic peptide AF17121 (Ac-VDECWRIIASHTWFCAEE) that inhibits interleukin 5 (IL-5) function and IL-5 receptor alpha-chain (IL-5Ralpha) binding has been derived from recombinant random peptide library screening and follow-up synthetic variation. To better understand the structural basis of its antagonist activity, AF17121 and a series of analogs of the parent peptide were prepared by solid phase peptide synthesis. Sequence variation was focused on the charged residues Asp(2), Glu(3), Arg(6), Glu(17), and Glu(18). Two of those residues, Glu(3) and Arg(6), form an EXXR motif that was found to be common among library-derived IL-5 antagonists. The E and R in the EXXR motif have a proximity similar to charged residues in a previously identified receptor alpha binding region, the beta-strand between the C- and D-helices of human IL-5. Optical biosensor interaction kinetics and cell proliferation assays were used to evaluate the antagonist activities of the purified synthetic peptides, by measuring competition with the highly active single chain IL-5. Analogs in which acidic residues (Asp(2), Glu(3), Glu(17), and Glu(18)) were replaced individually by Ala retained substantial competition activity, with multiple replacements in these residues leading to fractional loss of potency at most. In contrast, R6A analogs had strongly reduced competition activity. The results reveal that the arginine residue is crucial for the IL-5Ralpha binding of AF17121, while the acidic residues are not essential though likely complex-stabilizing particularly in the Asp(2)-Glu(3) region. By CD, AF17121 exhibited mostly disordered structure with evidence for a small beta-sheet content, and replacement of the arginine had no influence on the observed secondary structure of the peptides. The dominance of Arg(6) in AF17121 activity corresponds to previous findings of dominance of the positive charge balance in the antiparallel beta-sheet of IL-5 composed of (88)EERRR(92) in one strand of the CD turn region of IL-5 and with Arg(32) in the neighboring beta-strand. These results argue that AF17121 and related library-derived peptides function by mimicking the CD turn receptor alpha recognition epitope in IL-5 and open the way to small molecule antagonist design.

show abstract

Section: Introductionmentioning

confidence: 99%

Cyclic peptide interleukin 5 antagonists mimic CD turn recognition epitope for receptor α

et al. 2004

Self Cite

View full text Add to dashboard Cite

show abstract

“…A mimic of interleukin-5 (IL5) was identified by random mutagenesis of the turn in a coiled coil miniprotein (CCSL) using the phage display technique [32]. IL5 is a homodimer comprised of two -helical bundles [33].…”

Section: Receptor Bindingmentioning

confidence: 99%

Creating Functional Artificial Proteins

Razeghifard¹,

Wallace²,

Pace³

et al. 2007

CPPS

View full text Add to dashboard Cite

Much is now known about how protein folding occurs, through the sequence analysis of proteins of known folding geometry and the sequence/structural analysis of proteins and their mutants. This has allowed not only the modification of natural proteins but also the construction of de novo polypeptides with predictable folding patterns. Structure/function analysis of natural proteins is used to construct derived versions that retain a degree of biological activity. The constructed versions made of either natural or artificial sequences contain critical residues for activity such as receptor binding. In some cases, the functionality is introduced by incorporating binding sites for other elements, such as organic cofactors or transition metals, into the protein scaffold. While these modified proteins can mimic the function of natural proteins, they can also be constructed to have novel activities. Recently engineered photoactive proteins are good examples of such systems in which a light-induced electron transfer can be established in normally light-insensitive proteins. The present review covers some aspects of protein design that have been used to investigate protein receptor binding, cofactor binding and biological electron transfer.

show abstract

A survey of the year 2002 commercial optical biosensor literature

Rich

Myszka

2003

J of Molecular Recognition

View full text Add to dashboard Cite

We have compiled 819 articles published in the year 2002 that involved commercial optical biosensor technology. The literature demonstrates that the technology's application continues to increase as biosensors are contributing to diverse scientific fields and are used to examine interactions ranging in size from small molecules to whole cells. Also, the variety of available commercial biosensor platforms is increasing and the expertise of users is improving. In this review, we use the literature to focus on the basic types of biosensor experiments, including kinetics, equilibrium analysis, solution competition, active concentration determination and screening. In addition, using examples of particularly well-performed analyses, we illustrate the high information content available in the primary response data and emphasize the impact of including figures in publications to support the results of biosensor analyses.

show abstract

Coiled coil miniprotein randomization on phage leads to charge pattern mimicry of the receptor recognition determinant of interleukin 5

Cited by 6 publications

References 44 publications

Cyclic peptide interleukin 5 antagonists mimic CD turn recognition epitope for receptor α

Cyclic peptide interleukin 5 antagonists mimic CD turn recognition epitope for receptor α

Creating Functional Artificial Proteins

A survey of the year 2002 commercial optical biosensor literature

Contact Info

Product

Resources

About