Membrane asymmetry A survey and critical appraisal of the methodology II. Methods for assessing the unequal distribution of lipids

Etemadi, Abol-Hassan

doi:10.1016/0304-4157(80)90013-1

Cited by 4 publications

(5 citation statements)

References 348 publications

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“…Iodination of granules (3-5 mg/ml) or membranes (0.5-1 rag/ ml) in 250 mM sucrose, I mM EDTA, pH 7.4, was performed using Iodogen [14], as described by Fraker and Speck [15]. One rnl of suspension was added to a scintillation vial freshly coated with 10 ~tg of Iodogen, and 300 gCi of carrier-free Na…”

Section: Labeling Proceduresmentioning

confidence: 99%

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Isolation and partial characterization of platelet α-granule membranes

1983

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Porcine alpha-granules, prepared by a modification of pre-existing methods, were found to be essentially homogeneous by transmission electron microscopy. Freeze-fractured samples of isolated granules revealed numerous intramembranous particles on the EF (exoplasmic fracture) surface and to a lesser extent on the PF (protoplasmic fracture) surface whereas the PS (protoplasmic) surface was relatively smooth. The granules appear to be sealed, as evidenced by: a) the retention of their electron dense core material; b) the inability of impermeant labels to react with the granule contents, and c) the finding that the intragranular proteins are refractory to mild hydrolysis by externally added proteases. Membranes were isolated by alkali extraction of the granules and used for biochemical characterization. Approximately 87% of the protein, but only insignificant amounts of phospholipid were removed by this procedure, which yielded membrane vesicles devoid of the dense core. The membranes contain one major and several minor polypeptides of molecular weights ranging from 28,000 to 230,000, as determined by polyacrylamide gel electrophoresis. The major polypeptide contains carbohydrate residues. The exposure of specific proteins on the cytoplasmic surface of the granule membrane was determined by a combination of surface-specific labeling and proteolysis of intact granules, followed by membrane isolation and analysis. In sealed granules, only a limited number of bands are modified by the reagents whereas most of them are affected following granule lysis, indicating asymmetry in their transmembrane disposition. The fraction eluted by alkali extraction was also analyzed and found to contain nine major polypeptides of molecular weights ranging from 230,000 to 43,000. These are compared to the weights of the macromolecules believed to be secreted from alpha-granules, as determined by radioimmunological techniques.

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Section: Labeling Proceduresmentioning

confidence: 99%

“…Surface specific reagents were employed to investigate the exposure of g-granule membrane proteins to the cytoplasmic (external) milieu [14]. The granules were labeled by iodination using the sparingly soluble chloroamide Iodogen.…”

Section: Surface Labeling Of Intact Granulesmentioning

confidence: 99%

Isolation and partial characterization of platelet α-granule membranes

1983

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“…Even a complete loss of asymmetry in small subpopulations of a sample might not be detected using these methods. Using fluorescently labelled annexin V in flow cytometry was able to identify and quantify red cells that had lost their phospholipid asymmetry in subpopulations as small as 1 million cells .…”

Section: Introductionmentioning

confidence: 99%

The hypercoagulable status in common Mediterranean β‐thalassaemia mutations trait

Labib

Etewa

Atia

2014

Int J Lab Hematology

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“…Πραγματικά ? ύπάρχουν πληροφορίες οτι τά δομικά ένζυμα περιβάλλονται άπό ειδικά σέ κάθε περίπτωση φωσφολιπίδια (41,42). "Αλλο επιχείρημα πού συνηγορεί γιά τήν ασυμμετρία των λιπιδίων είνα.ι,οτι τό χημικό περι βάλλον κάθε πλευράς της μεμβράνης είναι διαφορετικό καί μπορεί ανάλογα νά προκαλεί καί νά διατηρεί τήν ει δική τοποθέτηση διαφόρων λιπιδίων.…”

Section: η μοριακή κίνηση και ρευστότητα της μεμβράνηςunclassified

Η Επιδραση Της Χοληστερολης Και Του Γλυκοζιδιου Της Επι Των Δραστικοτητων Μιτοχονδριακων Ενζυμων Εγκεφαλου Επιμυος

Κοπεικινα-Τσιμπουκιδου¹

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Γεννήθηκα στό Νόβο-Μίργοροντ της Ρωσσίας. Τελείω σα τό Κρατικό Πανεπιστήμιο Λομονοσοφ της Μόσχας -τή Σχολή της Βιολογίας -τό 1968. Πήρα τό πτυχίο Βιολόγου μέ ειδικότητα "Φυσιολογία" και ύποειδικότητα "Γενετική -Επιλογή", Στό τελευταίο έτος των σπουδών μου ασχολήθη κα ειδικά μέ τό θέμα : "Μελέτη μεταλλακτών E.coli Κ12 μέ διαταραχθείσα ικανότητα έπανασυνδέσεως". Τή μελέτη τοΰ θέματος αύτοϋ έκανα στην "Εδρα της Γενετικής και 'Επιλογής κάτω άπό τήν εποπτεία του Καθηγητή S.I.Alikhanian. Για τή σχετική εργασία μου ή Ειδική Κρατική 'Επι τροπή τοϋ Πανεπιστημίου μοΰ απένειμε τόν τίτλο "Master of Biology". Τό Νοέμβριο τοΰ 1968 ύστερα άπό εξετάσεις πήρα Κρατική υποτροφία γιά μετεκπαίδευση μου πάνω στην ειδι κότητα "Γενετική των Μικροβίων" και πάλι στην "Εδρα της Γενετικής και 'Επιλογής τοϋ Πανεπιστημίου της Μόσχας.

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Membrane asymmetry A survey and critical appraisal of the methodology II. Methods for assessing the unequal distribution of lipids

Cited by 4 publications

References 348 publications

Isolation and partial characterization of platelet α-granule membranes

Isolation and partial characterization of platelet α-granule membranes

The hypercoagulable status in common Mediterranean β‐thalassaemia mutations trait

Η Επιδραση Της Χοληστερολης Και Του Γλυκοζιδιου Της Επι Των Δραστικοτητων Μιτοχονδριακων Ενζυμων Εγκεφαλου Επιμυος

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