Folding transitions in calpain activator peptides studied by solution NMR spectroscopy

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Calpastatin, the endogenous inhibitor of calpain, a cysteine protease in eukaryotic cells, is an intrinsically unstructured protein, which upon binding to the enzyme goes through a conformational change. Peptides calpA (SGKSGMDAALDDLIDTLGG) and calpC (SKPIGPDDAIDALSSDFTS), corresponding to the two conserved subdomains of calpastatin, are known to activate calpain and increase the Ca2+ sensitivity of the enzyme. Using solution NMR spectroscopy, here we show that calpA and calpC are disordered in water but assume an α-helical conformation in 50% CD3OH.The position and length of the helices are in agreement with those described in the literature for the bound state of the corresponding segments of calpastatin suggesting that the latter might be structurally primed for the interaction with its target. According to our data, the presence of Ca2+ induces a backbone rearrangement in the peptides, an effect thatmay contribute to setting the fine conformational balance required for the interaction of the peptides with calpain.

Original languageEnglish
Pages (from-to)404-410
Number of pages7
JournalJournal of Peptide Science
Volume15
Issue number6
DOIs
Publication statusPublished - 2009

Fingerprint

Nuclear magnetic resonance spectroscopy
Calpain
Magnetic Resonance Spectroscopy
Peptides
Intrinsically Disordered Proteins
Cysteine Proteases
Eukaryotic Cells
Enzymes
Conformations
Water
calpain activator
calpastatin
calpain inhibitors

Keywords

  • Calcium-binding
  • Calpain
  • Calpastatin
  • Conformational change
  • Folding
  • Inhibitor
  • Intrinsically unstructured proteins
  • NMR spectroscopy
  • Order-disorder transition

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Medicine
  • Molecular Biology
  • Biochemistry
  • Pharmacology
  • Drug Discovery
  • Organic Chemistry
  • Medicine(all)

Cite this

@article{0bdadade6add42e8a338b55daf3c7f51,
title = "Folding transitions in calpain activator peptides studied by solution NMR spectroscopy",
abstract = "Calpastatin, the endogenous inhibitor of calpain, a cysteine protease in eukaryotic cells, is an intrinsically unstructured protein, which upon binding to the enzyme goes through a conformational change. Peptides calpA (SGKSGMDAALDDLIDTLGG) and calpC (SKPIGPDDAIDALSSDFTS), corresponding to the two conserved subdomains of calpastatin, are known to activate calpain and increase the Ca2+ sensitivity of the enzyme. Using solution NMR spectroscopy, here we show that calpA and calpC are disordered in water but assume an α-helical conformation in 50{\%} CD3OH.The position and length of the helices are in agreement with those described in the literature for the bound state of the corresponding segments of calpastatin suggesting that the latter might be structurally primed for the interaction with its target. According to our data, the presence of Ca2+ induces a backbone rearrangement in the peptides, an effect thatmay contribute to setting the fine conformational balance required for the interaction of the peptides with calpain.",
keywords = "Calcium-binding, Calpain, Calpastatin, Conformational change, Folding, Inhibitor, Intrinsically unstructured proteins, NMR spectroscopy, Order-disorder transition",
author = "Orsolya Toke and Z. B{\'a}n{\'o}czi and G. T{\'a}rk{\'a}nyi and P. Friedrich and F. Hudecz",
year = "2009",
doi = "10.1002/psc.1131",
language = "English",
volume = "15",
pages = "404--410",
journal = "Journal of Peptide Science",
issn = "1075-2617",
publisher = "John Wiley and Sons Ltd",
number = "6",

}

TY - JOUR

T1 - Folding transitions in calpain activator peptides studied by solution NMR spectroscopy

AU - Toke, Orsolya

AU - Bánóczi, Z.

AU - Tárkányi, G.

AU - Friedrich, P.

AU - Hudecz, F.

PY - 2009

Y1 - 2009

N2 - Calpastatin, the endogenous inhibitor of calpain, a cysteine protease in eukaryotic cells, is an intrinsically unstructured protein, which upon binding to the enzyme goes through a conformational change. Peptides calpA (SGKSGMDAALDDLIDTLGG) and calpC (SKPIGPDDAIDALSSDFTS), corresponding to the two conserved subdomains of calpastatin, are known to activate calpain and increase the Ca2+ sensitivity of the enzyme. Using solution NMR spectroscopy, here we show that calpA and calpC are disordered in water but assume an α-helical conformation in 50% CD3OH.The position and length of the helices are in agreement with those described in the literature for the bound state of the corresponding segments of calpastatin suggesting that the latter might be structurally primed for the interaction with its target. According to our data, the presence of Ca2+ induces a backbone rearrangement in the peptides, an effect thatmay contribute to setting the fine conformational balance required for the interaction of the peptides with calpain.

AB - Calpastatin, the endogenous inhibitor of calpain, a cysteine protease in eukaryotic cells, is an intrinsically unstructured protein, which upon binding to the enzyme goes through a conformational change. Peptides calpA (SGKSGMDAALDDLIDTLGG) and calpC (SKPIGPDDAIDALSSDFTS), corresponding to the two conserved subdomains of calpastatin, are known to activate calpain and increase the Ca2+ sensitivity of the enzyme. Using solution NMR spectroscopy, here we show that calpA and calpC are disordered in water but assume an α-helical conformation in 50% CD3OH.The position and length of the helices are in agreement with those described in the literature for the bound state of the corresponding segments of calpastatin suggesting that the latter might be structurally primed for the interaction with its target. According to our data, the presence of Ca2+ induces a backbone rearrangement in the peptides, an effect thatmay contribute to setting the fine conformational balance required for the interaction of the peptides with calpain.

KW - Calcium-binding

KW - Calpain

KW - Calpastatin

KW - Conformational change

KW - Folding

KW - Inhibitor

KW - Intrinsically unstructured proteins

KW - NMR spectroscopy

KW - Order-disorder transition

UR - http://www.scopus.com/inward/record.url?scp=67649743774&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67649743774&partnerID=8YFLogxK

U2 - 10.1002/psc.1131

DO - 10.1002/psc.1131

M3 - Article

VL - 15

SP - 404

EP - 410

JO - Journal of Peptide Science

JF - Journal of Peptide Science

SN - 1075-2617

IS - 6

ER -