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Antiapoptotic properties of ?-crystallin-derived peptide chaperones and characterization of their uptake transporters in human RPE cells.

ABSTRACT:

Purpose

The chaperone proteins, ?-crystallins, also possess antiapoptotic properties. The purpose of the present study was to investigate whether 19 to 20-mer ?-crystallin-derived mini-chaperone peptides (?-crystallin mini-chaperone) are antiapoptotic, and to identify their putative transporters in human fetal RPE (hfRPE) cells.

Methods

Cell death and caspase-3 activation induced by oxidative stress were quantified in early passage hfRPE cells in the presence of 19 to 20-mer ?A- or ?B-crystallin-derived or scrambled peptides. Cellular uptake of fluorescein-labeled, ?-crystallin-derived mini-peptides and recombinant full-length ?B-crystallin was determined in confluent hfRPE. The entry mechanism in hfRPE cells for ?-crystallin mini-peptides was investigated. The protective role of polycaprolactone (PCL) nanoparticle encapsulated ?B-crystallin mini-chaperone peptides from H2O2-induced cell death was studied.

Results

Primary hfRPE cells exposed to oxidative stress and either ?A- or ?B-crystallin mini-chaperones remained viable and showed marked inhibition of both cell death and activation of caspase-3. Uptake of full-length ?B-crystallin was minimal while a time-dependent uptake of ?B-crystallin-derived peptide was observed. The mini-peptides entered the hfRPE cells via the sodium-coupled oligopeptide transporters 1 and 2 (SOPT1, SOPT2). PCL nanoparticles containing ?B-crystallin mini-chaperone were also taken up and protected hfRPE from H2O2-induced cell death at significantly lower concentrations than free ?B-crystallin mini-chaperone peptide.

Conclusions

?A- and ?B-crystallin mini-chaperones offer protection to hfRPE cells and inhibit caspase-3 activation. The oligopeptide transporters SOPT1 and SOPT2 mediate the uptake of these peptides in RPE cells. Nanodelivery of ?B-crystallin-derived mini-chaperone peptide offers an alternative approach for protection of hfRPE cells from oxidant injury.

SUBMITTER: Sreekumar PG

PROVIDER: S-EPMC3632268 | biostudies-literature | 2013 Apr

REPOSITORIES: biostudies-literature

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Publications

Antiapoptotic properties of α-crystallin-derived peptide chaperones and characterization of their uptake transporters in human RPE cells.

Sreekumar Parameswaran G PG Chothe Paresh P Sharma Krishna K KK Baid Rinku R Kompella Uday U Spee Christine C Kannan Nandini N Manh Christina C Ryan Stephen J SJ Ganapathy Vadivel V Kannan Ram R Hinton David R DR

Investigative ophthalmology & visual science 20130417 4

<h4>Purpose</h4>The chaperone proteins, α-crystallins, also possess antiapoptotic properties. The purpose of the present study was to investigate whether 19 to 20-mer α-crystallin-derived mini-chaperone peptides (α-crystallin mini-chaperone) are antiapoptotic, and to identify their putative transporters in human fetal RPE (hfRPE) cells.<h4>Methods</h4>Cell death and caspase-3 activation induced by oxidative stress were quantified in early passage hfRPE cells in the presence of 19 to 20-mer αA- o ...[more]

PMID: 23532520

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Project description:The demonstration of chaperone-like activity in peptides (mini-chaperones) derived from ?-crystallin's chaperone region has generated significant interest in exploring the therapeutic potential of peptide chaperones in diseases of protein aggregation. Recent studies in experimental animals show that mini-chaperones could reach intended targets and alter the disease phenotype. Although mini-chaperones show potential benefits against protein aggregation diseases, they do tend to form aggregates on storage. There is thus a need to fine-tune peptide chaperones to increase their solubility, pharmacokinetics, and biological efficacy.This review summarizes the properties and the potential therapeutic roles of mini-chaperones in protein aggregation diseases and highlights some of the refinements needed to increase the stability and biological efficacy of mini-chaperones while maintaining or enhancing their chaperone-like activity against precipitation of unfolding proteins.Mini-chaperones suppress the aggregation of proteins, block amyloid fibril formation, stabilize mutant proteins, sequester metal ions, and exhibit antiapoptotic properties. Much work must be done to fine-tune mini-chaperones and increase their stability and biological efficacy. Peptide chaperones could have a great therapeutic value in diseases associated with protein aggregation and apoptosis.Accumulation of misfolded proteins is a primary cause for many age-related diseases, including cataract, macular degeneration, and various neurological diseases. Stabilization of native proteins is a logical therapeutic approach for such diseases. Mini-chaperones, with their inherent antiaggregation and antiapoptotic properties, may represent an effective therapeutic molecule to prevent the cascade of protein conformational disorders. Future studies will further uncover the therapeutic potential of mini-chaperones. This article is part of a Special Issue entitled Crystallin Biochemistry in Health and Disease.