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The bovine seminal plasma protein PDC-109 extracts phosphorylcholine-containing lipids from the outer membrane leaflet

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Abstract

The bovine seminal plasma protein PDC-109 modulates the maturation of bull sperm cells by removing lipids, mainly phosphatidylcholine and cholesterol, from their cellular membrane. Here, we have characterized the process of extraction of endogenous phospholipids and of their respective analogues. By measuring the PDC-109-mediated release of fluorescent phospholipid analogues from lipid vesicles and from biological membranes (human erythrocytes, bovine epididymal sperm cells), we showed that PDC-109 extracts phospholipids with a phosphorylcholine headgroup mainly from the outer leaflet of these membranes. The ability of PDC-109 to extract endogenous phospholipids from epididymal sperm cells was followed by mass spectrometry, which allowed us to characterize the fatty acid pattern of the released lipids. From these cells, PDC-109 extracted phosphatidylcholine and sphingomyelin that contained an enrichment of mono- and di-unsaturated fatty acids as well as short-chain and lyso-phosphatidylcholine species. Based on the results, a model explaining the phospholipid specificity of PDC-109-mediated lipid release is presented.

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Abbreviations

BSA:

Bovine serum albumin

HBS:

Hepes buffered salt solution

L/P:

Lipid to protein molar ratio

LUV:

Large unilamellar vesicle(s)

MALDI-TOF MS:

Matrix-assisted laser desorption and ionization time-of-flight mass spectrometry

NBD-PC:

1-Palmitoyl-2-[6-[(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]caproyl]phosphatidylcholine

NBD-PE:

1-Palmitoyl-2-[6-[(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]caproyl]phosphatidylethanolamine

NBD-PL:

NBD-labelled phospholipids

NBD-PS:

1-palmitoyl-2-[6-[(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]caproyl]phosphatidylserine

NBD-SM:

6-[(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]hexanoyl)sphingosylphosphatidylcholine

PC:

Phosphatidylcholine

PE:

Phosphatidylethanolamine

PS:

Phosphatidylserine

RBC:

Red blood cells

SM:

Sphingomyelin

TLC:

Thin layer chromatography

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Acknowledgment

We are grateful to Mrs. Sabine Schiller (Humboldt University) for technical assistance. This work was supported by grants from the Deutsche Forschungsgemeinschaft to KM (Mu 1520/2-1), JS (Schi 476/5-1), ETP (To 114/4) and to PM (Mu 1017/2).

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Authors and Affiliations

  1. Institute of Biology, Humboldt University of Berlin, Invalidenstrasse 42, 10115, Berlin, Germany

    Astrid Tannert, Anke Kurz, Karl-Rudolf Erlemann, Andreas Herrmann & Peter Müller

  2. Institute for Reproduction of Farm Animals Schönow e.V., Bernauer Chaussee 10, 16321, Schönow, Germany

    Anke Kurz & Karin Müller

  3. Medical Faculty, Institute of Medical Physics and Biophysics, University of Leipzig, 04103, Leipzig, Germany

    Jürgen Schiller

  4. School of Veterinary Medicine Hannover, Institute of Reproductive Medicine, Bünteweg 15, 30559, Hannover, Germany

    Edda Töpfer-Petersen

  5. Department of Medicine, University of Montreal and Guy-Bernier Research Centre, Maisonneuve-Rosemont Hospital, H1T 2M4, Montreal, QC, Canada

    Puttaswamy Manjunath

  6. Department of Pharmacology, Charité-Universitätsmedizin Berlin, Thielallee 67-73, 14195, Berlin, Germany

    Astrid Tannert

  7. Charles River Laboratories, CTBR, Preclinical Services, 87 Chemin Senneville, H9X 3R3, Senneville (Montréal), QC, Canada

    Karl-Rudolf Erlemann

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  1. Astrid Tannert
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  2. Anke Kurz
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  4. Karin Müller
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  5. Andreas Herrmann
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  7. Edda Töpfer-Petersen
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  8. Puttaswamy Manjunath
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  9. Peter Müller
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Correspondence to Peter Müller.

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Astrid Tannert and Anke Kurz have contributed equally to this work.

Dedicated to Prof. K. Arnold on the occasion of his 65th birthday.

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Tannert, A., Kurz, A., Erlemann, KR. et al. The bovine seminal plasma protein PDC-109 extracts phosphorylcholine-containing lipids from the outer membrane leaflet. Eur Biophys J 36, 461–475 (2007). https://doi.org/10.1007/s00249-006-0105-3

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