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Monolayer–multilayer transitions in a lung surfactant model: IR reflection–absorption spectroscopy and atomic force microscopy

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Abstract

A hydrophobic pulmonary surfactant protein, SP-C, has been implicated in surface-associated activities thought to facilitate the work of breathing. Model surfactant films composed of lipids and SP-C display a reversible transition from a monolayer to surface-associated multilayers upon compression and expansion at the air/water (A/W) interface. The molecular-level mechanics of this process are not yet fully understood. The current work uses atomic force microscopy on Langmuir–Blodgett films to verify the formation of multilayers in a dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylglycerol, cholesterol, and SP-C model system. Isotherms of SP-C-containing films are consistent with exclusion and essentially complete respreading during compression and expansion, respectively. Multilayer formation was not detected in the absence of SP-C. Most notable are the results from IR reflection–absorption spectroscopy (IRRAS) conducted at the A/W interface, where the position and intensity of the Amide I band of SP-C reveal that the predominantly helical structure changes its orientation in monolayers versus multilayers. IRRAS measurements indicate that the helix tilt angle changed from approximately 80° in monolayers to a transmembrane orientation in multilayers. The results constitute the first quantitative measure of helix orientation in mixed monolayer/multilamellar domains at the A/W interface and provide insight into the molecular mechanism for SP-C-facilitated respreading of surfactant.

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Abbreviations

AFM:

Atomic force microscopy

ATR:

Attenuated total reflectance

A/W:

Air/water

DLigPC:

1,2-Dilignoceroyl phosphatidylcholine

DPPC:

1,2-Dipalmitoylphosphatidylcholine

DPPG:

1,2-Dipalmitoylphosphatidylglycerol

DSPC:

1,2-Distearoylphosphatidylcholine

EDTA:

Ethylenediaminetetraacetate

IRRAS:

IR reflection–absorption spectroscopy

LB:

Langmuir–Blodgett

PC:

Phosphatidylcholine

PG:

Phosphatidylglycerol

π−A:

Surface pressure–molecular area

POPG:

1-Palmitoyl-2-oleoyl phosphatidylglycerol

SFM:

Scanning force microscopy

TOF-SIMS:

Time-of-flight secondary ion mass spectrometry

Tris:

Tris(hydroxymethyl)aminomethane

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Acknowledgements

This work was supported by PHS grant GM 29864 to R.M.

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Authors and Affiliations

  1. Department of Chemistry, Newark College of Arts and Sciences, Rutgers University, 73 Warren Street, Newark, NJ, 07102, USA

    Lin Wang, Peng Cai, Huixin He, Carol R. Flach & Richard Mendelsohn

  2. Institut für Biochemie, Westfälische Wilhelms-Universität, 48149, Münster, Germany

    Hans-Joachim Galla

Authors
  1. Lin Wang
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  2. Peng Cai
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  3. Hans-Joachim Galla
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  4. Huixin He
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  5. Carol R. Flach
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  6. Richard Mendelsohn
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Correspondence to Carol R. Flach.

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Wang, L., Cai, P., Galla, HJ. et al. Monolayer–multilayer transitions in a lung surfactant model: IR reflection–absorption spectroscopy and atomic force microscopy. Eur Biophys J 34, 243–254 (2005). https://doi.org/10.1007/s00249-004-0446-8

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  • DOI: https://doi.org/10.1007/s00249-004-0446-8

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