Isolation of plasma membranes from mammary gland by two-phase polymer partitioning

Keenan, T.W.; Valivullah, H. Mohammed; Dunlevy, Jane T.

doi:10.1016/0003-2697(89)90039-0

Cited by 11 publications

(5 citation statements)

References 15 publications

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“…The sheet-like plasma membranes were low in relative activities of marker enzymes and protein yield (0.185 mg per g wet tissue). This was greater than the yield of 0.032 mg obtained from plasma membranes isolated from lactating cow mammary gland, but less than that of isolated PM from lactating rat mammary gland by two-phase polymer partitioning (0.3 ± 0.1 mg) . The marker enzymes used in the present study, 5′-nucleotidase and Na + /K + -ATPase, are considered to be reliable as PM markers of lactating mammary cells .…”

Section: Discussioncontrasting

confidence: 51%

“…This was greater than the yield of 0.032 mg obtained from plasma membranes isolated from lactating cow mammary gland, 12 but less than that of isolated PM from lactating rat mammary gland by two-phase polymer partitioning (0.3 ± 0.1 mg). 5 The marker enzymes used in the present study, 5′-nucleotidase and Na + /K + -ATPase, are considered to be reliable as PM markers of lactating mammary cells. 12 Their activities in the PM were only 9 or 11 times greater than in the homogenates; perhaps the contamination of cytoplasmic proteins originating from the outer mitochondrial membranes caused this result.…”

Section: ■ Discussionmentioning

confidence: 87%

“…Fortunately, tandem mass spectrometry is currently the most powerful tool for PM proteome analysis on the condition that the PM fraction has been efficiently purified. Differential centrifugation combined with density gradient ultracentrifugation and two-phase aqueous partition 5 are traditional methods to enrich the PM. Immunoisolation, biotin−avidin, and agglutinin-affinity purification have also been used to enrich the PM from rat liver, 6 cancer cells, 7 and prostate cells, 8 respectively.…”

Section: ■ Introductionmentioning

confidence: 99%

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Proteomic Analysis of Isolated Plasma Membrane Fractions from the Mammary Gland in Lactating Cows

Yan

Tang

Tan

et al. 2015

J. Agric. Food Chem.

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The mammary gland of dairy cows is a formidable lipid-synthesizing machine for lactation. This unique function depends on the activities of plasma membrane (PM) proteins in mammary cells. Little information is known about the expression profiles of PM proteins and their functions during the lactating process. This study investigated the proteome map of PM fractions of mammary gland in lactating cows using 1D-Gel-LC-MS/MS and identified 872 nonredundant proteins with 141 unknown proteins, wherein 215 were PM-associated proteins. Most of the PM-associated proteins were binding, transport, and catalytic proteins such as annexin proteins, heat shock proteins, integrins, RAS oncogene family members, and S100 calcium binding proteins. The PM-associated pathways such as caveolae-mediated endocytosis, leukocyte extravasation, aldosterone signaling in epithelial cells, and remodeling of epithelial adherens junctions were also significantly over-represented. Proteomic analysis revealed the characteristics and predicted functions of PM proteins isolated from the lactating bovine mammary gland. These results further provide experimental evidence for the presence of many proteins predicted in the annotated bovine genome. The data generated here also provide a reference for the PM-related functional research in the mammary gland of lactating cows.

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Section: Discussioncontrasting

confidence: 51%

Section: ■ Discussionmentioning

confidence: 87%

Section: ■ Introductionmentioning

confidence: 99%

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Proteomic Analysis of Isolated Plasma Membrane Fractions from the Mammary Gland in Lactating Cows

Yan

Tang

Tan

et al. 2015

J. Agric. Food Chem.

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“…MLGM was suspended in the detergent-containing solution by using an all-glass homogenizer, and held on ice for 30 to 60 min, after which insoluble and soluble materials were separated by centrifugation for 60 min at 2 mC and 150 000 g. Mammary tissues were collected from rats between the seventh and fourteenth days of lactation, and from lactating cows slaughtered at a meat-packing plant. Endoplasmic reticulum [6], cytoplasmic lipid droplet [5], cytosol [6], plasma membrane [13] and Golgi apparatus [14] fractions were isolated from tissue homogenates. Epididymal fat pads were collected from male Sprague-Dawley rats of 150 g body weight, and adipocytes were prepared from this tissue by dissociation with collagenase [15].…”

Section: Membranes and Cell Fractionsmentioning

confidence: 99%

Adipocyte differentiation-related protein is secreted into milk as a constituent of milk lipid globule membrane

Heid

Schnölzer

Keenan

1996

Biochemical Journal

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131

133

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Milk lipid globules from humans, cows and rats contained a protein identified as adipocyte differentiation-related protein (ADRP) associated with the globule surface membrane material. This protein, previously believed to be specific to adipocytes, was a major constituent of the globule surface and was present in a detergent-insoluble complex that contained stoichiometric amounts of butyrophilin and xanthine oxidase. Identification of ADRP was by sequence similarity of tryptic peptides from cow and human proteins with the sequence inferred from the cDNA for mouse ADRP. The putative ADRP of lipid globules from cow, human and rat milk was recognized specifically by antisera raised against a peptide synthesized to duplicate the N-terminal 26 residues of the mouse protein. In homogenates of lactating mammary gland, ADRP was found only in endoplasmic reticulum and in lipid droplet fractions. ADRP was modified, apparently post-translationally, and one modification apparently was acylation, primarily with C14, C16 and C18 fatty acids. Two isoelectric variants of ADRP were present in cow globule membrane material. In vitro, ADRP served as a substrate for protein kinases associated with milk lipid globule membrane, but this protein did not seem to become phosphorylated intracellularly.

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“…Non-specific binding by the biotinylated egg PM was always removed prior to its use as a probe by exposure to liver PM proteins. Porcine liver cell PM was isolated (Keenan et al, 1989), solubilised in 0.1% CHAPS and preadsorbed to a sheet of Immobilon-P (Millipore, Bedford, MA) for 6 h. The sheet of Immobilon-P was blocked overnight in Tween Trisbuffered saline (TTBS; 100 mM Tris, 0.9% NaCl, 0.1% v/v Tween 20, pH 7.4) at 4°C. The egg PM probe was incubated with this blocked sheet of Immobilon-P for 1 h prior to probing the sperm PM blots.…”

Section: Egg Pm Isolation and Biotinylationmentioning

confidence: 99%

Identification of porcine sperm plasma membrane proteins that may play a role in sperm–egg fusion

Ash

Berger

Horner

et al. 1995

Zygote

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SummarySperm plasma membrane (PM) proteins that demonstrate affinity for egg PM preparations have the potential to be biologically important during sperm-egg binding and/or fusion. In this study four such proteins have been identified. To provide quantitative evidence for possible biological function, the large natural variation among different porcine sperm populations with regard to their ability to interact with the egg was compared with the relative binding of egg PM material to individual proteins. An aliquot from each of 24 porcine ejaculates was evaluated by the zona-free hamster ova bioassay and the remainder processed to yield sperm PM vesicles. Aliquots of sperm PM were solubilised, separated by SDS-PAGE, western blotted and probed with partially purified, biotinylated egg PM protein. Bound egg PM proteins were visualised on western blots by an avidin/biotin/horseradish peroxidase system and analysed by scanning laser densitometry. Four sperm PM proteins (62, 39, 27 and 7 kDa estimated molecular mass) were the predominant binders of egg PM. The amount of egg PM bound to the 62 kDa protein was significantly correlated with the ability of sperm from the 24 ejaculates to penetrate zona-free hamster ova (percentage of ova penetrated, p = 0.01, R = 0.65; number of penetrated sperm per ovum, p = 0.02, R = 0.63)

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Isolation of plasma membranes from mammary gland by two-phase polymer partitioning

Cited by 11 publications

References 15 publications

Proteomic Analysis of Isolated Plasma Membrane Fractions from the Mammary Gland in Lactating Cows

Proteomic Analysis of Isolated Plasma Membrane Fractions from the Mammary Gland in Lactating Cows

Adipocyte differentiation-related protein is secreted into milk as a constituent of milk lipid globule membrane

Identification of porcine sperm plasma membrane proteins that may play a role in sperm–egg fusion

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