Characterization of the in situ structural and interfacial properties of the cationic hydrophobic heteropolypeptide, KL4, in lung surfactant bilayer and monolayer models at the air - Water interface: Implications for pulmonary surfactant delivery

Heidi - Mansour, Srinivasan Damodaran, George Zografi

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

This study examines the various equilibrium in situ secondary structures of the pharmaceutical heteropolypeptide, KL4, in the solid state, in solution, and in the monolayer state alone and mixed with dipalmitoylphosphatidylcholine (DPPC) and palmitoyloleoylphosphatidylglycerol (POPG). In situ surface circular dichroism spectroscopy, using a method first reported by Damodaran (Damodaran, S. Anal. Bioanal. Chem. 2003, 376, 182-188), of equilibrated KL4, DPPC/KL4, POPG/KL4, and DPPC/POPG/KL4 monolayers at the air - water interface was used to examine the in situ two-dimensional conformation of KL4. Gravimetric vapor sorption by solid KL4 was used to analyze the effects of water molecules on the conformation of KL4 when confined as a monolayer at the surface of water. Solid-state KL4 conformation was determined by X-ray powder diffraction (XRPD). The equilibrium interfacial and spreading properties were measured at 25 °C, 37 °C, and 45 °C using the Wilhelmy plate method and Langmuir film balance. Equilibrium phase transition temperatures were measured using differential scanning calorimetry (DSC). It was found that solid-state KL4, which takes up very little water, exhibits β-sheet and α-helix secondary structures, whereas KL4 in solution appears to exist only as an α-helix. KL4 forms a stable, insoluble monolayer, exhibiting β-sheet and aperiodic structures. These structures provide KL4, when confined in two-dimensions, the structural flexibility to maximize favorable cationic lysine-water interactions and favorable leucine - leucine hydrophobic and van der Waals interactions; while effectively "shielding" the leucine residues away from water. In DPPC/KL4 monolayers, KL4 retains its native β-sheet and aperiodic structures, consistent with phase separation of DPPC and KL4, in bilayers and monolayers. In POPG/KL4 monolayers, KL4 exhibits an increase in aperiodic secondary structures (loss of β-sheet) to maximize favorable electrostatic interactions, consistent with the observed negative deviations from ideal monolayer mixing.

Original languageEnglish (US)
Pages (from-to)681-695
Number of pages15
JournalMolecular Pharmaceutics
Volume5
Issue number5
DOIs
StatePublished - Sep 2008
Externally publishedYes

Fingerprint

Pulmonary Surfactants
1,2-Dipalmitoylphosphatidylcholine
Surface-Active Agents
Air
Lung
Water
Leucine
Powder Diffraction
Transition Temperature
Phase Transition
Differential Scanning Calorimetry
Circular Dichroism
Static Electricity
X-Ray Diffraction
Lysine
Spectrum Analysis
Pharmaceutical Preparations

Keywords

  • Bilayer - monolayer equilibria
  • Biomacromolecule conformation/secondary structure
  • In situ surface circular dichroism
  • Langmuir films
  • Molecular interactions
  • Phase transitions
  • Phospholipids
  • Solid-state
  • Surface structure
  • Water vapor absorption/hydration

ASJC Scopus subject areas

  • Pharmaceutical Science
  • Molecular Medicine
  • Drug Discovery

Cite this

@article{afea57619d734e56b5a7be7be5426dec,
title = "Characterization of the in situ structural and interfacial properties of the cationic hydrophobic heteropolypeptide, KL4, in lung surfactant bilayer and monolayer models at the air - Water interface: Implications for pulmonary surfactant delivery",
abstract = "This study examines the various equilibrium in situ secondary structures of the pharmaceutical heteropolypeptide, KL4, in the solid state, in solution, and in the monolayer state alone and mixed with dipalmitoylphosphatidylcholine (DPPC) and palmitoyloleoylphosphatidylglycerol (POPG). In situ surface circular dichroism spectroscopy, using a method first reported by Damodaran (Damodaran, S. Anal. Bioanal. Chem. 2003, 376, 182-188), of equilibrated KL4, DPPC/KL4, POPG/KL4, and DPPC/POPG/KL4 monolayers at the air - water interface was used to examine the in situ two-dimensional conformation of KL4. Gravimetric vapor sorption by solid KL4 was used to analyze the effects of water molecules on the conformation of KL4 when confined as a monolayer at the surface of water. Solid-state KL4 conformation was determined by X-ray powder diffraction (XRPD). The equilibrium interfacial and spreading properties were measured at 25 °C, 37 °C, and 45 °C using the Wilhelmy plate method and Langmuir film balance. Equilibrium phase transition temperatures were measured using differential scanning calorimetry (DSC). It was found that solid-state KL4, which takes up very little water, exhibits β-sheet and α-helix secondary structures, whereas KL4 in solution appears to exist only as an α-helix. KL4 forms a stable, insoluble monolayer, exhibiting β-sheet and aperiodic structures. These structures provide KL4, when confined in two-dimensions, the structural flexibility to maximize favorable cationic lysine-water interactions and favorable leucine - leucine hydrophobic and van der Waals interactions; while effectively {"}shielding{"} the leucine residues away from water. In DPPC/KL4 monolayers, KL4 retains its native β-sheet and aperiodic structures, consistent with phase separation of DPPC and KL4, in bilayers and monolayers. In POPG/KL4 monolayers, KL4 exhibits an increase in aperiodic secondary structures (loss of β-sheet) to maximize favorable electrostatic interactions, consistent with the observed negative deviations from ideal monolayer mixing.",
keywords = "Bilayer - monolayer equilibria, Biomacromolecule conformation/secondary structure, In situ surface circular dichroism, Langmuir films, Molecular interactions, Phase transitions, Phospholipids, Solid-state, Surface structure, Water vapor absorption/hydration",
author = "Mansour, {Heidi -} and Srinivasan Damodaran and George Zografi",
year = "2008",
month = "9",
doi = "10.1021/mp700123p",
language = "English (US)",
volume = "5",
pages = "681--695",
journal = "Molecular Pharmaceutics",
issn = "1543-8384",
publisher = "American Chemical Society",
number = "5",

}

TY - JOUR

T1 - Characterization of the in situ structural and interfacial properties of the cationic hydrophobic heteropolypeptide, KL4, in lung surfactant bilayer and monolayer models at the air - Water interface

T2 - Implications for pulmonary surfactant delivery

AU - Mansour, Heidi -

AU - Damodaran, Srinivasan

AU - Zografi, George

PY - 2008/9

Y1 - 2008/9

N2 - This study examines the various equilibrium in situ secondary structures of the pharmaceutical heteropolypeptide, KL4, in the solid state, in solution, and in the monolayer state alone and mixed with dipalmitoylphosphatidylcholine (DPPC) and palmitoyloleoylphosphatidylglycerol (POPG). In situ surface circular dichroism spectroscopy, using a method first reported by Damodaran (Damodaran, S. Anal. Bioanal. Chem. 2003, 376, 182-188), of equilibrated KL4, DPPC/KL4, POPG/KL4, and DPPC/POPG/KL4 monolayers at the air - water interface was used to examine the in situ two-dimensional conformation of KL4. Gravimetric vapor sorption by solid KL4 was used to analyze the effects of water molecules on the conformation of KL4 when confined as a monolayer at the surface of water. Solid-state KL4 conformation was determined by X-ray powder diffraction (XRPD). The equilibrium interfacial and spreading properties were measured at 25 °C, 37 °C, and 45 °C using the Wilhelmy plate method and Langmuir film balance. Equilibrium phase transition temperatures were measured using differential scanning calorimetry (DSC). It was found that solid-state KL4, which takes up very little water, exhibits β-sheet and α-helix secondary structures, whereas KL4 in solution appears to exist only as an α-helix. KL4 forms a stable, insoluble monolayer, exhibiting β-sheet and aperiodic structures. These structures provide KL4, when confined in two-dimensions, the structural flexibility to maximize favorable cationic lysine-water interactions and favorable leucine - leucine hydrophobic and van der Waals interactions; while effectively "shielding" the leucine residues away from water. In DPPC/KL4 monolayers, KL4 retains its native β-sheet and aperiodic structures, consistent with phase separation of DPPC and KL4, in bilayers and monolayers. In POPG/KL4 monolayers, KL4 exhibits an increase in aperiodic secondary structures (loss of β-sheet) to maximize favorable electrostatic interactions, consistent with the observed negative deviations from ideal monolayer mixing.

AB - This study examines the various equilibrium in situ secondary structures of the pharmaceutical heteropolypeptide, KL4, in the solid state, in solution, and in the monolayer state alone and mixed with dipalmitoylphosphatidylcholine (DPPC) and palmitoyloleoylphosphatidylglycerol (POPG). In situ surface circular dichroism spectroscopy, using a method first reported by Damodaran (Damodaran, S. Anal. Bioanal. Chem. 2003, 376, 182-188), of equilibrated KL4, DPPC/KL4, POPG/KL4, and DPPC/POPG/KL4 monolayers at the air - water interface was used to examine the in situ two-dimensional conformation of KL4. Gravimetric vapor sorption by solid KL4 was used to analyze the effects of water molecules on the conformation of KL4 when confined as a monolayer at the surface of water. Solid-state KL4 conformation was determined by X-ray powder diffraction (XRPD). The equilibrium interfacial and spreading properties were measured at 25 °C, 37 °C, and 45 °C using the Wilhelmy plate method and Langmuir film balance. Equilibrium phase transition temperatures were measured using differential scanning calorimetry (DSC). It was found that solid-state KL4, which takes up very little water, exhibits β-sheet and α-helix secondary structures, whereas KL4 in solution appears to exist only as an α-helix. KL4 forms a stable, insoluble monolayer, exhibiting β-sheet and aperiodic structures. These structures provide KL4, when confined in two-dimensions, the structural flexibility to maximize favorable cationic lysine-water interactions and favorable leucine - leucine hydrophobic and van der Waals interactions; while effectively "shielding" the leucine residues away from water. In DPPC/KL4 monolayers, KL4 retains its native β-sheet and aperiodic structures, consistent with phase separation of DPPC and KL4, in bilayers and monolayers. In POPG/KL4 monolayers, KL4 exhibits an increase in aperiodic secondary structures (loss of β-sheet) to maximize favorable electrostatic interactions, consistent with the observed negative deviations from ideal monolayer mixing.

KW - Bilayer - monolayer equilibria

KW - Biomacromolecule conformation/secondary structure

KW - In situ surface circular dichroism

KW - Langmuir films

KW - Molecular interactions

KW - Phase transitions

KW - Phospholipids

KW - Solid-state

KW - Surface structure

KW - Water vapor absorption/hydration

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U2 - 10.1021/mp700123p

DO - 10.1021/mp700123p

M3 - Article

C2 - 18630875

AN - SCOPUS:56049099316

VL - 5

SP - 681

EP - 695

JO - Molecular Pharmaceutics

JF - Molecular Pharmaceutics

SN - 1543-8384

IS - 5

ER -