Direct Enthalpy Measurements of the Calcium-Dependent Interaction of Annexin V and VI with Phospholipid Vesicles

Biochemistry ◽  
1994 ◽  
Vol 33 (45) ◽  
pp. 13239-13249 ◽  
Author(s):  
Douglas A Plager ◽  
Gary L. Nelsestuen
Keyword(s):  
Annexin V ◽  
Phospholipid Vesicles ◽  
Calcium Dependent ◽  
Dependent Interaction

Annexin V forms calcium-dependent trimeric units on phospholipid vesicles

FEBS Letters ◽  
1992 ◽  
Vol 314 (2) ◽  
pp. 159-162 ◽  
Author(s):  
Nestor O. Concha ◽  
James F. Head ◽  
Marcia A. Kaetzel ◽  
John R. Dedman ◽  
Barbara A. Seaton
Keyword(s):  
Annexin V ◽  
Phospholipid Vesicles ◽  
Calcium Dependent

The Calcium-Dependent Binding of Annexin V to Phospholipid Vesicles Influences the Bilayer Inner Fluidity Gradient†

Biochemistry ◽  
1998 ◽  
Vol 37 (29) ◽  
pp. 10540-10546 ◽  
Author(s):  
Francesco M. Megli ◽  
Mariangela Selvaggi ◽  
Susanne Liemann ◽  
Ernesto Quagliariello ◽  
Robert Huber
Keyword(s):  
Annexin V ◽  
Phospholipid Vesicles ◽  
Calcium Dependent

Glycoproteins IIb and IIIa and Platelet Thrombospondin in a Liposome Model of Platelet Aggregation

1986 ◽  
Vol 55 (02) ◽  
pp. 240-245 ◽  
Author(s):  
M E Rybak
Keyword(s):  
Platelet Aggregation ◽  
Direct Evidence ◽  
Phospholipid Vesicles ◽  
Aggregate Formation ◽  
Membrane Glycoproteins ◽  
Protein Orientation ◽  
Phosphatidylcholine Liposomes ◽  
Calcium Dependent ◽  
Lentil Lectin ◽  
Complete Reversal

SummaryPlatelet membrane glycoproteins IIb and IIIa and platelet thrombospondin were incorporated onto phosphatidylcholine liposomes, by freeze thawing and sonication. Protein orientation on the liposomes was confirmed by susceptibility to neuraminidase cleavage and binding to lentil lectin-Sepharose (GPIIb-IIIa liposomes) and to heparin-Sepharose (thrombospondin liposomes). Glycoproteins Ilb-IIIa bound 125I-fibrinogen with Kd of 7.5 × 10™7M. Binding was reversible and calcium-dependent. Ilb-IIIa liposomes underwent fibrinogen-dependent aggregation in the presence of 10 mM CaCl2. Maximal aggregate formation was observed with a combination of IIb-IIIa liposomes and thrombospondin liposomes. This aggregation was partially inhibited by preincubation with monoclonal antibodies to the IIb-IIIa complex. Addition of EDTA caused complete reversal of aggregates. Thrombospondin liposomes also underwent fibrinogen and calcium dependent aggregation, however, this aggregation was less than that observed with the GPIIb-IIIa liposomes. Maximal aggregate formation was observed with a mixture of IIb-IIIa liposomes and thrombospondin liposomes. These studies demonstrate that GPIIb-IIIa and thrombospondin can be incorporated into phospholipid vesicles with preservation of function. Direct evidence is provided to demonstrate that glycoprotein lib and Ilia and fibrinogen are sufficient for platelet aggregation and to demonstrate that thrombospondin may also contribute to platelet aggregation.

scholarly journals Isolation and characterization of two distinct low-density, Triton-insoluble, complexes from porcine lung membranes

1996 ◽  
Vol 319 (3) ◽  
pp. 887-896 ◽  
Author(s):  
Edward T PARKIN ◽  
Anthony J TURNER ◽  
Nigel M HOOPER
Keyword(s):  
Light Scattering ◽  
Gradient Centrifugation ◽  
Buoyant Density ◽  
Annexin V ◽  
Density Fraction ◽  
Calcium Dependent ◽  
Membrane Complex ◽  
Isolation And Characterization ◽  
Insoluble Complex ◽  
Marker Proteins

The Triton-insoluble complex from porcine lung membranes has been separated into two distinct subfractions visible as discrete light-scattering bands following buoyant density-gradient centrifugation in sucrose. Both of these detergent-insoluble complexes were enriched in the glycosyl-phosphatidylinositol (GPI)-anchored ectoenzymes alkaline phosphatase, aminopeptidase P and 5´-nucleotidase, and both complexes excluded the polypeptide-anchored ectoenzymes angiotensin-converting enzyme, dipeptidyl peptidase IV and aminopeptidases A and N. The GPI-anchored proteins in both complexes were susceptible to release by phosphatidylinositol-specific phospholipase C. Both complexes were also enriched in cholesterol and glycosphingolipids, and in caveolin/VIP21, although only the higher-density fraction was enriched in the plasmalemmal caveolar marker proteins Ca2+-ATPase and the inositol 1,4,5-trisphosphate receptor. Among the annexin family of proteins, annexins I and IV were absent from the two detergent-insoluble complexes, annexin V was present in both, and annexins II and VI were only enriched in the higher-density fraction. When the metal chelator EGTA was present in the isolation buffers, annexins II and VI dissociated from the higher-density detergent-insoluble complex and only a single light-scattering band was observed on the sucrose gradient, at the same position as for the lower-density complex. In contrast, in the presence of excess calcium only a single detergent-insoluble complex was isolated from the sucrose gradients, at an intermediate density. Thus the detergent-insoluble membrane complex can be subfractionated on the basis of what appears to be calcium-dependent, annexin-mediated, vesicle aggregation into two distinct populations, only one of which is enriched in plasmalemmal caveolar marker proteins.

Monoclonal antibodies against human α-fetoprotein exploitation of an unusual calcium-dependent interaction with the antigen for analytical and preparative purposes

1988 ◽  
Vol 111 (1) ◽  
pp. 67-73 ◽  
Author(s):  
Irena Štefanová ◽  
Václav Hořejší ◽  
Hana Krištofová ◽  
Pavla Angelisová ◽  
Václav Žižkovský ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Calcium Dependent ◽  
Dependent Interaction

scholarly journals Selective Ca2+-dependent interaction of calmodulin with the head domain of synapsin 1

1991 ◽  
Vol 275 (1) ◽  
pp. 93-97 ◽  
Author(s):  
N V Hayes ◽  
A F Bennett ◽  
A J Baines
Keyword(s):  
Regulatory Protein ◽  
Critical Element ◽  
Nerve Terminals ◽  
Calcium Dependent ◽  
Calmodulin Binding ◽  
Head Domain ◽  
N Terminus ◽  
Dependent Interaction ◽  
Synapsin 1

The calcium-dependent regulatory protein calmodulin is a critical element in the machinery regulating exocytosis at nerve terminals. Okabe & Sobue [(1987) FEBS Lett. 213, 184-188] showed that calmodulin interacts with one of the proteins intimately connected with the neuronal exocytotic process, i.e. synapsin 1. We have investigated the site at which calmodulin interacts with synapsin 1. We find that it is possible to generate chemically cross-linked Ca2(+)-dependent complexes between synapsin 1 and calmodulin in vitro, and have used covalent cross-linking in conjunction with calmodulin affinity chromatography to identify fragments of synapsin 1 that interact with calmodulin. Ca2(+)-dependent calmodulin binding is restricted to the ‘head’ domain (residues 1-453 in bovine synapsin 1). Within this domain the binding site is located in a unique 11 kDa Staphylococcus aureus V8 proteinase generated fragment. This fragment does not contain the site for cyclic-AMP-dependent phosphorylation and therefore does not represent the N-terminus of the protein.

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