The protein and lipid composition of the membrane of milk fat globules depends on their size

Jing Lu, Nurit Argov-Argaman, Jeni Anggrek, Sjef Boeren, Toon van Hooijdonk, Jacques Vervoort, Kasper Arthur Hettinga

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Abstract

In bovine milk, fat globules (MFG) have a heterogeneous size distribution with diameters ranging from 0.1 to 15 μm. Although efforts have been made to explain differences in lipid composition, little is known about the protein composition of MFG membranes (MFGM) in different sizes of MFG. In this study, protein and lipid analyses were combined to study MFG formation and secretion. Two different sized MFG fractions (7.6 ± 0.9 μm and 3.3 ± 1.2 μm) were obtained by centrifugation. The protein composition of MFGM in the large and small MFG fractions was compared using mass-spectrometry-based proteomics techniques. The lipid composition and fatty acid composition of MFG was determined using HPLC-evaporative light-scattering detector and gas chromatography, respectively. Two frequently studied proteins in lipid droplet biogenesis, perilipin-2 and TIP47, were increased in the large and small MFG fractions, respectively. In the large MFG fraction, besides perilipin-2, cytoplasmic vesicle proteins (heat shock proteins, 14-3-3 proteins, and Rabs), microfilaments and intermediate filament-related proteins (actin and vimentin), host defense proteins (cathelicidins), and phosphatidylinositol were higher in concentration. On the other hand, cholesterol synthesis enzymes [lanosterol synthase and sterol-4-α-carboxylate 3-dehydrogenase (decarboxylating)], cholesterol, unsaturated fatty acids, and phosphatidylethanolamine were, besides TIP47, higher in concentration in the small MFG fraction. These results suggest that vesicle proteins, microfilaments and intermediate filaments, cholesterol, and specific phospholipids play an important role in lipid droplet growth, secretion, or both. The observations from this study clearly demonstrated the difference in protein and lipid composition between small and large MFG fractions. Studying the role of these components in more detail in future experiments may lead to a better understanding of fat globule formation and secretion.

LanguageEnglish
Article number74498
Pages4726-4738
JournalJournal of Dairy Science
Volume99
Issue number6
DOIs
Publication statusPublished - 2016

Fingerprint

fat globules
protein composition
Membrane Lipids
lipid composition
milk fat
Proteins
adipophilin
14-3-3 Proteins
Lipids
Intermediate Filament Proteins
intermediate filaments
proteins
cholesterol
secretion
Cathelicidins
microfilament proteins
droplets
Fats
Cholesterol
milk fat globule

Keywords

  • Fat globule
  • Membrane proteome
  • Milk
  • Phospholipids
  • Triglycerides

Cite this

Lu, Jing ; Argov-Argaman, Nurit ; Anggrek, Jeni ; Boeren, Sjef ; van Hooijdonk, Toon ; Vervoort, Jacques ; Hettinga, Kasper Arthur. / The protein and lipid composition of the membrane of milk fat globules depends on their size. In: Journal of Dairy Science. 2016 ; Vol. 99, No. 6. pp. 4726-4738.
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abstract = "In bovine milk, fat globules (MFG) have a heterogeneous size distribution with diameters ranging from 0.1 to 15 μm. Although efforts have been made to explain differences in lipid composition, little is known about the protein composition of MFG membranes (MFGM) in different sizes of MFG. In this study, protein and lipid analyses were combined to study MFG formation and secretion. Two different sized MFG fractions (7.6 ± 0.9 μm and 3.3 ± 1.2 μm) were obtained by centrifugation. The protein composition of MFGM in the large and small MFG fractions was compared using mass-spectrometry-based proteomics techniques. The lipid composition and fatty acid composition of MFG was determined using HPLC-evaporative light-scattering detector and gas chromatography, respectively. Two frequently studied proteins in lipid droplet biogenesis, perilipin-2 and TIP47, were increased in the large and small MFG fractions, respectively. In the large MFG fraction, besides perilipin-2, cytoplasmic vesicle proteins (heat shock proteins, 14-3-3 proteins, and Rabs), microfilaments and intermediate filament-related proteins (actin and vimentin), host defense proteins (cathelicidins), and phosphatidylinositol were higher in concentration. On the other hand, cholesterol synthesis enzymes [lanosterol synthase and sterol-4-α-carboxylate 3-dehydrogenase (decarboxylating)], cholesterol, unsaturated fatty acids, and phosphatidylethanolamine were, besides TIP47, higher in concentration in the small MFG fraction. These results suggest that vesicle proteins, microfilaments and intermediate filaments, cholesterol, and specific phospholipids play an important role in lipid droplet growth, secretion, or both. The observations from this study clearly demonstrated the difference in protein and lipid composition between small and large MFG fractions. Studying the role of these components in more detail in future experiments may lead to a better understanding of fat globule formation and secretion.",
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The protein and lipid composition of the membrane of milk fat globules depends on their size. / Lu, Jing; Argov-Argaman, Nurit; Anggrek, Jeni; Boeren, Sjef; van Hooijdonk, Toon; Vervoort, Jacques; Hettinga, Kasper Arthur.

In: Journal of Dairy Science, Vol. 99, No. 6, 74498, 2016, p. 4726-4738.

Research output: Contribution to journalArticleAcademicpeer-review

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T1 - The protein and lipid composition of the membrane of milk fat globules depends on their size

AU - Lu, Jing

AU - Argov-Argaman, Nurit

AU - Anggrek, Jeni

AU - Boeren, Sjef

AU - van Hooijdonk, Toon

AU - Vervoort, Jacques

AU - Hettinga, Kasper Arthur

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AB - In bovine milk, fat globules (MFG) have a heterogeneous size distribution with diameters ranging from 0.1 to 15 μm. Although efforts have been made to explain differences in lipid composition, little is known about the protein composition of MFG membranes (MFGM) in different sizes of MFG. In this study, protein and lipid analyses were combined to study MFG formation and secretion. Two different sized MFG fractions (7.6 ± 0.9 μm and 3.3 ± 1.2 μm) were obtained by centrifugation. The protein composition of MFGM in the large and small MFG fractions was compared using mass-spectrometry-based proteomics techniques. The lipid composition and fatty acid composition of MFG was determined using HPLC-evaporative light-scattering detector and gas chromatography, respectively. Two frequently studied proteins in lipid droplet biogenesis, perilipin-2 and TIP47, were increased in the large and small MFG fractions, respectively. In the large MFG fraction, besides perilipin-2, cytoplasmic vesicle proteins (heat shock proteins, 14-3-3 proteins, and Rabs), microfilaments and intermediate filament-related proteins (actin and vimentin), host defense proteins (cathelicidins), and phosphatidylinositol were higher in concentration. On the other hand, cholesterol synthesis enzymes [lanosterol synthase and sterol-4-α-carboxylate 3-dehydrogenase (decarboxylating)], cholesterol, unsaturated fatty acids, and phosphatidylethanolamine were, besides TIP47, higher in concentration in the small MFG fraction. These results suggest that vesicle proteins, microfilaments and intermediate filaments, cholesterol, and specific phospholipids play an important role in lipid droplet growth, secretion, or both. The observations from this study clearly demonstrated the difference in protein and lipid composition between small and large MFG fractions. Studying the role of these components in more detail in future experiments may lead to a better understanding of fat globule formation and secretion.

KW - Fat globule

KW - Membrane proteome

KW - Milk

KW - Phospholipids

KW - Triglycerides

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DO - 10.3168/jds.2015-10375

M3 - Article

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