Structural Characterization of Disulfide-Linked p53-Derived Peptide Dimers

DiGiorno, Magdalena C.; Vithanage, Nisansala; Victorio, Clara G.; Kreitler, Dale F.; Outlaw, Victor K.; Sawyer, Nicholas

doi:10.1007/s10989-024-10651-5

Title: Structural Characterization of Disulfide-Linked p53-Derived Peptide Dimers

Journal Article · 01 October 2024 · International journal of peptide research and therapeutics (Online)

DOI: https://doi.org/10.1007/s10989-024-10651-5 · OSTI ID:2477954

DiGiorno, Magdalena C. ^[1]; Vithanage, Nisansala ^[2]; Victorio, Clara G. ^[1];

^[3];

^[2];

^[1]

Fordham Univ., The Bronx, NY (United States)
Univ. of Missouri, Columbia, MO (United States)
Brookhaven National Laboratory (BNL), Upton, NY (United States). National Synchrotron Light Source II (NSLS-II)

Disulfide bonds provide a convenient method for chemoselective alteration of peptide and protein structure and function. We previously reported that mild oxidation of a p53-derived bisthiol peptide (CTFANLWRLLAQNC) under dilute non-denaturing conditions led to unexpected disulfide-linked dimers as the exclusive product. The dimers were antiparallel, significantly α-helical, resistant to protease degradation, and easily reduced back to the original bisthiol peptide. Here, in this work, we examine the intrinsic factors influencing peptide dimerization using a combination of amino acid substitution, circular dichroism (CD) spectroscopy, and X-ray crystallography. CD analysis of peptide variants suggests critical roles for Leu6 and Leu10 in the formation of stable disulfide-linked dimers. The 1.0 Å resolution crystal structure of the peptide dimer supports these data, revealing a leucine-rich LxxLL dimer interface with canonical knobs-into-holes packing. Two levels of higher-order oligomerization are also observed in the crystal: an antiparallel “dimer of dimers” mediated by Phe3 and Trp7 residues in the asymmetric unit and a tetramer of dimers mediated by Trp7 and Leu10. In CD spectra of Trp-containing peptide variants, minima at 227 nm provide evidence for the dimer of dimers in dilute aqueous solution. Importantly, and in contrast to the original dimer model, the canonical leucine-rich core and robust dimerization of most peptide variants suggests a tunable molecular architecture to target various proteins and evaluate how folding and oligomerization impact various properties, such as cell permeability.

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Research Organization:: Brookhaven National Laboratory (BNL), Upton, NY (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities (SUF); USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0012704

OSTI ID:: 2477954

Report Number(s):: BNL--226344-2024-JAAM

Journal Information:: International journal of peptide research and therapeutics (Online), Journal Name: International journal of peptide research and therapeutics (Online) Journal Issue: 6 Vol. 30; ISSN 1573-3904

Publisher:: Springer NatureCopyright Statement

Country of Publication:: United States

Language:: English

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36 MATERIALS SCIENCE
dimerization
disulfide
folding
structure

Title: Structural Characterization of Disulfide-Linked p53-Derived Peptide Dimers

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