6,862 research outputs found
Recruitment of AP-1 clathrin adaptors to liposomal membranes
No full textProtein and membrane traffic between organelles within the endocytic and exocytic pathway is mediated most prominently by coated vesicles. These vesicles are formed by the assembly of cytosolic coat proteins onto the donor membrane, which deform it into a bud so that vesicles can pinch off. Clathrin with its associated adaptors, COPI and COPII are the three major coats. Various in vitro studies allowed insight into the mechanism of coat formation. COPI and COPII vesicle budding from chemically defined liposomes has been reconstituted in vitro, using pure coat compounds. Further, it has been demonstrated that cargo is sorted into these vesicles. The mechanism of clathrin-coated vesicle formation appears to be more complicated. The AP-1 clathrin adaptor is involved in vesicle formation at the transGolgi network and endosomes. This work presents an in vitro assay where AP-1 is recruited to peptidoliposomes, presenting covalently linked peptides corresponding to sorting signals. In this system, AP-1 recruitment depends on myristyolated ADP-ribosylation factor(ARF1), GTP or GMP-PNP, tyrosine signals and a small amount of phosphoinositides, most prominently phosphatidyl inositol 4,5bisphosphate. In such a minimal system AP-1 is recruited as a highmolecular weight complex indicating the formation of a precoat in the absence of clathrin. GTP hydrolysis, induced by ARF GTPase-activating protein(ARFGAP1), disassembled this complex. Further, AP-1 is able to enhance the GAP activity of ARFGAP1 on myristoylated ARF1, suggesting a regulatory function of GTP hydrolysis in early steps of coat recruitment. This work provides insights into the mechanism of AP-1 clathrin coat formation which might also be used to investigate the recruitment of other coats
A role for amphiphysin in AP-1/clathrin coat formation
No full textTransport of cargo within the endocytic and secretory pathway is generally mediated by coated vesicles. Clathrin, in combination with different adaptor proteins, is the major coat protein for vesicle formation at the plasma membrane, endosomes, and the trans-Golgi network (TGN). Best characterized is the formation of clathrin coats for endocytosis at the plasma membrane involving the adaptor protein complex AP-2. Clathrin and AP-2 were shown to be at the centre of a complex interactome of proteins accessory to vesicle formation. Considerably less is known about the formation of clathrin coated carriers at the TGN and endosomes, where the adaptor protein complex AP-1 plays a major role.
In vitro studies showed the minimal requirements for association of AP-1 to liposomal membranes to be activated ARF1, phosphoinositides, and either sorting signals or unknown cytosolic factors. We have used a liposome floatation assay to identify cytosolic proteins collaborating with AP-1 at the membrane. Separation of proteins from bovine brain cytosol with several chromatographic methods yielded an active fraction containing amphiphysin 1, amphiphysin 2, and endophilin A1. All three proteins are expressed in brain and known to be involved in AP-2/clathrin coat formation. They consist of an N-terminal N-BAR (Bin, amphiphysin, Rvs) domain for dimerization and membrane binding and a C-terminal SH3 (Src homology 3) domain for interaction with dynamin and synaptojanin. Amphiphysin 1 and 2 in addition contain a middle domain with binding sites for adaptors and clathrin. It was proposed that amphiphysins and endophilin are targeted to membranes with high curvature, such as the neck of a forming vesicle, where they recruit dynamin and synaptojanin in preparation for vesicle fission and uncoating.
In this thesis, I bacterially expressed and purified all three proteins and tested them in the floatation assay for AP-1 membrane binding activity. Only amphiphysin 2 showed activity, both as a homodimer and as a heterodimer with amphiphysin 1. Activity depended on a motif that was shown to bind to AP-1, AP-2, and clathrin in GST pull-down experiments.
Endogenous amphiphysins in primary neurons, as well as transiently expressed in neuronal or fibroblast cell lines, co-localized with AP-1 at the TGN. In addition, when expressed at high levels in neuronal cells, amphiphysins aggregated and interfered dominantly with the TGN localization of AP-1. Both phenomena depended on the presence of the clathrin and adaptor interaction sequence in the amphiphysins. Furthermore, both amphiphysins could be cross-linked to AP-1 in vivo.
Our results indicate that amphiphysin 1 and 2 function not only in clathrin coated vesicle formation for endocytosis at the plasma membrane, but are also part of the machinery forming AP-1/clathrin coats at the TGN and endosomes. This suggests that the machineries for CCV formation with AP-1 and AP-2 at different locations in the cell share more components than previously anticipated
A cytosolic factor mediating membrane recruitment of AP-1 clathrin adaptors
No full textTransport of cargo within the endocytic and secretory pathway is generally mediated by coated vesicles. These vesicles are formed through the recruitment of cytosolic coat proteins to the donor membrane that act as a scaffold to form coated buds and vesicles. At the same time they selectively concentrate cargo proteins by interacting with cytosolic signals. Clathrin, in combination with different adaptor proteins (APs), is the major coat protein for vesicle formation at the plasma membrane, endosomes and the trans-Golgi network. Best characterized is clathrin mediated endocytosis at the plasma membrane which involves AP-2 and a network of associated proteins. Much less is known about AP-1 mediated clathrin coated vesicle formation at the TGN/endosomes. In vitro studies demonstrated that the minimal requirements to recruit AP-1 to liposome membranes are activated Arf1, phosphoinositides, and either sorting signals or an unknown cytosolic factor. In order to identify this factor, we fractionated calf brain cytosol by several chromatographic methods. Fractions were tested for factor dependent AP-1 recruitment activity using an in vitro assay. Purification via ammonium sulfate precipitation, hydrophobic interaction chromatography, anion/cation exchange chromatography or hydroxyapatite chromatography produced a final fraction containing three major proteins: amphiphysin 1, amphiphysinand endophilin A1. All three proteins are known accessory factors in clathrin coated vesicle formation at the plasma membrane. Co-immunodepletion of amphiphysinandresulted in a strong reduction of AP-1 recruitment activity. Therefore we conclude that a heterodimer of amphiphysinandis the long searched for cytosolic factor, required to recruit AP-1 in the absence of sorting signals in vitro. Our results strongly suggest that amphiphysin 1, amphiphysinand endophilin A1 are also involved in AP-1 mediated clathrin coated vesicle formation at the TGN and endosomes in vivo
John Robert Townshend, 1st Earl Sydney - Statesmen No. 13
No full textApeBildbeschriftung: "334", "VANITY FAIR.", "No. 26 STATESMEN, No. 13", "Price 6d.", "He received the Royal Commands and lengthened the skirts of the Ballet.", "May 1, 1869."Signatur: "Ape" = Carlo Pellegrini.Bleistiftvermerk: "Viscount Sidney".Das Magazin "Vanity Fair" wurde von Thomas Gibson Bowles veröffentlicht.Siehe auch: https://www.npg.org.uk/collections/search/portrait/mw255607/John-Robert-Townshend-1st-Earl-Sydney-Statesmen-No-13?set=523%3BVanity+Fair+cartoons%3A+chromolithographs+1869-1914&search=ap&rNo=1
The Treatment of Ties in AP Correlation
No full textThe Kendall tau and AP correlation coefficients are very commonly use to compare two rankings over the same set of items. Even though Kendall tau was originally defined assuming that there are no ties in the rankings, two alternative versions were soon developed to account for ties in two different scenarios: measure the accuracy of an observer with respect to a true and objective ranking, and measure the agreement between two observers in the absence of a true ranking. These two variants prove useful in cases where ties are possible in either ranking, and may indeed result in very different scores. AP correlation was devised to incorporate a top-heaviness component into Kendall tau, penalizing more heavily if differences occur between items at the top of the rankings, making it a very compelling coefficient in Information Retrieval settings. However, the treatment of ties in AP correlation remains an open problem. In this paper we fill this gap, providing closed analytical formulations of AP correlation under the two scenarios of ties contemplated in Kendall tau. In addition,we developed an R package that implements these coefficients.Best Short Paper Accepted author manuscriptMultimedia ComputingWeb Information System
John Robert Townshend, 1st Earl Sydney - Statesmen No. 13
No full textApeBildbeschriftung: "334", "VANITY FAIR.", "No. 26 STATESMEN, No. 13", "Price 6d.", "He received the Royal Commands and lengthened the skirts of the Ballet.", "May 1, 1869."Signatur: "Ape" = Carlo Pellegrini.Bleistiftvermerk: "Viscount Sidney".Das Magazin "Vanity Fair" wurde von Thomas Gibson Bowles veröffentlicht.Siehe auch: https://www.npg.org.uk/collections/search/portrait/mw255607/John-Robert-Townshend-1st-Earl-Sydney-Statesmen-No-13?set=523%3BVanity+Fair+cartoons%3A+chromolithographs+1869-1914&search=ap&rNo=1
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Irrigation's Impact on Society
No full textThe Anthropological Papers of the University of Arizona is a peer-reviewed monograph series sponsored by the School of Anthropology. Established in 1959, the series publishes archaeological and ethnographic papers that use contemporary method and theory to investigate problems of anthropological importance in the southwestern United States, Mexico, and related areas.List of Illustrations / Preface - Theodore E. Downing and McGuire Gibson / Historic Patterns of Mesopotamian Irrigation Agriculture - Robert McC. Adams / Violation of Fallow and Engineered Disaster in Mesopotamian Civilization - McGuire Gibson / Sassanian and Early Islamic Water-Control and Irrigation Systems on the Deh Luran Plain, Iran - James A. Neely / Irrigation and Society: The Iranian Plateau - Brian Spooner / Water Control in Terraced Rice-Field Agriculture in Southeastern Asia - J. E. Spencer / The System Nobody Knows: Village Irrigation in the Swiss Alps - Robert McC. Netting / Organizational Preadaptation to Irrigation: The Evolution of Early Water-Management Systems in Coastal Peru - M. Edward Moseley / Irrigation and Settlement Pattern: Preliminary Research Results from the North Coast of Peru - Ian Farrington / Conservation and Diversion: Water-Control Systems in the Anasazi Southwest - R. Gwinn Vivian / Irrigation and Moisture-Sensitive Periods: A Zapotec Case - Theodore E. Downing / The State's Use of Irrigation in Changing Peasant Society - Susan H. Lees / Irrigation, Conflict, and Politics: A Mexican Case - Eva Hunt and Robert C. Hunt / Irrigation Development and Population Pressure - Wayne Kappel / Comments on the Symposium From a Hydrologic Perspective - Martin M. Fogel / Comments on the Symposium From a Geographic Perspective - Chris Field / IndexThis title from the Anthropological Papers of the University of Arizona collection is made available by the University of Arizona Press and University of Arizona Libraries. If you have questions about this title, please contact the UA Press at http://www.uapress.arizona.edu/
Delamination Analysis of A Class of AP-PLY Composite Laminates
No full textA recently developed fiber placement architecture, AP-PLY, has been shown to give significantly improved damage tolerance characteristics of composite structures. The behavior of delaminations resulting from low speed impact damage is of particular concern. Major attention has been paid to expand current knowledge on the delamination response of simple AP-PLY composite structure and move towards in-depth understanding of the failure mechanisms behind the damage tolerance. This thesis presents the approaches to predict delamination onset and analyze delamination growth, in support of the search of the optimum woven pattern for AP-PLY composite laminates. The recovered interlaminar stress between layers combined with the maximum stress criterion determined the delamination onset of simple AP-PLY composite laminate under out-of-plane loads. 2D finite element models with cohesive elements inserted in the interfaces of woven layers have been built to evaluate the delamination initiation and propagation in the different woven patterns of simple AP-PLY composite beams. The parameters of the woven pattern, such as the woven angle, the number of woven plies, the number of straight filled plies, and the location of the woven patterns in through the thickness direction, were investigated and shown to have a significant effect on delamination creation and growth. An energy method based on beam theory was proposed to analyze the strain energy release rate (SERR) of an existing crack in an AP-PLY beam structure. The developed analytical method was implemented in isotropic materials and the obtained SERR of a crack was validated by reference results and finite element solutions. The general behavior of crack growth on the left or right crack tip was evaluated and basic trends leading to crack propagation on one side of the crack were established. A correction factor was introduced to improve the accuracy of the SERR of a small crack through the numerical calculation. The singularity of crack tip caused by dissimilar materials was investigated and was found that the inclusion of the singularity effect could increase the accuracy for small cracks. It has been shown that the neutral axis needs to be relocated to decouple the bending and membrane behavior of unsymmetrical composite laminates, thus to meet the requirement of minimizing the strain energy of the delaminated beam to calculate the SERR of a delaminated composite beam. The calculated SERR of a crack in a composite beam has been verified by comparing with a finite element model. The woven plies in AP-PLY composite laminate altered the layup and two conventional laminates with different stacking sequences were identified in an AP-PLY composite laminate based on the assumption that the resin areas were ignored. A step by step approach was developed to obtain the SERR of a crack that goes across different materials. The analytical SERR determined when two materials are used in sequence, sets the stage for optimization of AP-PLY composite laminates without taking account of the effect of the resin area. The procedure of optimization of simple AP-PLY pattern was proposed and industry may benefit for many applications. An equivalent stiffness approach was used to model regions containing resin pockets and straight or inclined composite layers. A series of three point bending tests was carried out where the failure process and loading capacity were evaluated. The methodology, procedure of optimization, philosophy outlined in this thesis might also be applied to the more complicated fully woven AP-PLY composite laminates. The work in this thesis contributes to the understanding of the behavior of AP-PLY composite laminates with delaminations
Structure function analysis of blazars AP Librae and 3c279
No full textHighest Honors in AstronomyBlazars AP Librae and 3c279 are analyzed for microvariability using a technique known as structure function analysis. AP Librae was observed in April
of 2005 and 3c279 was observed in April of 2007. The data for AP Librae was
previously reduced by Andrew Collazzi and the author reduced the data for
3c279. Both sets of data were reduced using Robert Knop's data reduction
program.
The author ran structure function analysis on both sets of data. Structure function analysis is a statistical analysis run on data that is suppose to
nd timescales of variability, periodicity, and the noise type of data. Previous analysis of AP Librae confirmed mircrovariability, which also shows up in
the structure function of AP Librae. Blazar 3c279 was much quieter than AP
Librae and showed no microvariability durning any of the nights.College of Arts and ScienceDepartment of Physics and Astronom
AP-based wireless intrusion detection systems
No full textThis thesis was scanned from the print manuscript for digital preservation and is copyright the author.
Researchers can access this thesis by asking their local university, institution or public library to
make a request on their behalf. Monash staff and postgraduate students can use the link in the References field
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