pKa Determination of a Histidine Residue in a Short Peptide Using Raman Spectroscopy

Brett H. Pogostin, Anders Malmendal, Casey H. Londergan, Karin S. Åkerfeldt

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Determining the pKa of key functional groups is critical to understanding the pH-dependent behavior of biological proteins and peptide-based biomaterials. Traditionally, 1H NMR spectroscopy has been used to determine the pKa of amino acids; however, for larger molecules and aggregating systems, this method can be practically impossible. Previous studies concluded that the C-D stretches in Raman are a useful alternative for determining the pKa of histidine residues. In this study, we report on the Raman application of the C2-D probe on histidine's imidazole side chain to determining the pKa of histidine in a short peptide sequence. The pKa of the tripeptide was found via difference Raman spectroscopy to be 6.82, and this value was independently confirmed via 1H NMR spectroscopy on the same peptide. The C2-D probe was also compared to other Raman reporters of the protonation state of histidine and was determined to be more sensitive and reliable than other protonation-dependent signals. The C2-D Raman probe expands the tool box available to chemists interested in directly interrogating the pKa's of histidine-containing peptide and protein systems.
OriginalsprogEngelsk
ArtikelnummerE405
TidsskriftMolecules
Vol/bind24
Udgave nummer3
ISSN1420-3049
DOI
StatusUdgivet - 2019
Udgivet eksterntJa

Emneord

  • 1 H NMR spectroscopy
  • Acid dissociation constant
  • Deuterium replacement
  • Histidine
  • Isotopic labeling
  • Peptides
  • Proteins
  • Raman spectroscopy
  • Vibrational probes

Citer dette

Pogostin, Brett H. ; Malmendal, Anders ; Londergan, Casey H. ; Åkerfeldt, Karin S. / pKa Determination of a Histidine Residue in a Short Peptide Using Raman Spectroscopy. I: Molecules. 2019 ; Bind 24, Nr. 3.
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title = "pKa Determination of a Histidine Residue in a Short Peptide Using Raman Spectroscopy",
abstract = "Determining the pKa of key functional groups is critical to understanding the pH-dependent behavior of biological proteins and peptide-based biomaterials. Traditionally, 1H NMR spectroscopy has been used to determine the pKa of amino acids; however, for larger molecules and aggregating systems, this method can be practically impossible. Previous studies concluded that the C-D stretches in Raman are a useful alternative for determining the pKa of histidine residues. In this study, we report on the Raman application of the C2-D probe on histidine's imidazole side chain to determining the pKa of histidine in a short peptide sequence. The pKa of the tripeptide was found via difference Raman spectroscopy to be 6.82, and this value was independently confirmed via 1H NMR spectroscopy on the same peptide. The C2-D probe was also compared to other Raman reporters of the protonation state of histidine and was determined to be more sensitive and reliable than other protonation-dependent signals. The C2-D Raman probe expands the tool box available to chemists interested in directly interrogating the pKa's of histidine-containing peptide and protein systems.",
keywords = "1 H NMR spectroscopy, Acid dissociation constant, Deuterium replacement, Histidine, Isotopic labeling, Peptides, Proteins, Raman spectroscopy, Vibrational probes",
author = "Pogostin, {Brett H.} and Anders Malmendal and Londergan, {Casey H.} and {\AA}kerfeldt, {Karin S.}",
year = "2019",
doi = "10.3390/molecules24030405",
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pKa Determination of a Histidine Residue in a Short Peptide Using Raman Spectroscopy. / Pogostin, Brett H.; Malmendal, Anders; Londergan, Casey H.; Åkerfeldt, Karin S.

I: Molecules, Bind 24, Nr. 3, E405, 2019.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - pKa Determination of a Histidine Residue in a Short Peptide Using Raman Spectroscopy

AU - Pogostin, Brett H.

AU - Malmendal, Anders

AU - Londergan, Casey H.

AU - Åkerfeldt, Karin S.

PY - 2019

Y1 - 2019

N2 - Determining the pKa of key functional groups is critical to understanding the pH-dependent behavior of biological proteins and peptide-based biomaterials. Traditionally, 1H NMR spectroscopy has been used to determine the pKa of amino acids; however, for larger molecules and aggregating systems, this method can be practically impossible. Previous studies concluded that the C-D stretches in Raman are a useful alternative for determining the pKa of histidine residues. In this study, we report on the Raman application of the C2-D probe on histidine's imidazole side chain to determining the pKa of histidine in a short peptide sequence. The pKa of the tripeptide was found via difference Raman spectroscopy to be 6.82, and this value was independently confirmed via 1H NMR spectroscopy on the same peptide. The C2-D probe was also compared to other Raman reporters of the protonation state of histidine and was determined to be more sensitive and reliable than other protonation-dependent signals. The C2-D Raman probe expands the tool box available to chemists interested in directly interrogating the pKa's of histidine-containing peptide and protein systems.

AB - Determining the pKa of key functional groups is critical to understanding the pH-dependent behavior of biological proteins and peptide-based biomaterials. Traditionally, 1H NMR spectroscopy has been used to determine the pKa of amino acids; however, for larger molecules and aggregating systems, this method can be practically impossible. Previous studies concluded that the C-D stretches in Raman are a useful alternative for determining the pKa of histidine residues. In this study, we report on the Raman application of the C2-D probe on histidine's imidazole side chain to determining the pKa of histidine in a short peptide sequence. The pKa of the tripeptide was found via difference Raman spectroscopy to be 6.82, and this value was independently confirmed via 1H NMR spectroscopy on the same peptide. The C2-D probe was also compared to other Raman reporters of the protonation state of histidine and was determined to be more sensitive and reliable than other protonation-dependent signals. The C2-D Raman probe expands the tool box available to chemists interested in directly interrogating the pKa's of histidine-containing peptide and protein systems.

KW - 1 H NMR spectroscopy

KW - Acid dissociation constant

KW - Deuterium replacement

KW - Histidine

KW - Isotopic labeling

KW - Peptides

KW - Proteins

KW - Raman spectroscopy

KW - Vibrational probes

U2 - 10.3390/molecules24030405

DO - 10.3390/molecules24030405

M3 - Journal article

VL - 24

JO - Molecules

JF - Molecules

SN - 1420-3049

IS - 3

M1 - E405

ER -