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COP Coated Vesicles00:59

COP Coated Vesicles

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Membrane-enclosed structures called vesicles transport proteins and lipids across the cell. The vesicles derive their cargo from the plasma membrane, Golgi, ER, or endosome. Coated vesicles are spherical, protein-coated carriers with a 50–100 nm diameter that mediate bidirectional transport between the ER and the Golgi. The distribution of proteins between the ER and Golgi complex is dynamic and is maintained by different coated vesicles. Their formation is driven by the assembly of...
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Pinching-off of Coated Vesicles01:32

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Vesicle budding is orchestrated by distinct cytosolic proteins such as adaptor proteins, coat proteins, and GTPases. To initiate vesicle budding, membrane-bending proteins containing crescent-shaped BAR domains bind to the lipid heads in the bilayer and distort the membrane to form a protein-coated vesicle bud. Adaptors proteins such as AP2 for clathrin-coated vesicles can nucleate on the deformed membrane. Finally, coat proteins such as clathrin or COPI and COPII assemble into a coat forming...
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Coat Assembly and GTPases01:33

Coat Assembly and GTPases

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Vesicles incorporate different coat protein subunits in different cell locations, which changes the properties of the coat, such as the shape and geometry of the transport vesicles. Thus, vesicle coat proteins also play a significant role in cargo selection.
Coat assembly depends on the local availability of phosphatidylinositol phosphates or PIPs and GTP-binding proteins. Adaptor proteins, which link the coat proteins to the membrane, bind to these PIPs and play a crucial role in controlling...
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Vesicular Tubular Clusters01:45

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After budding out from the ER membrane, some COPII vesicles lose their coat and fuse with one another to form larger vesicles and interconnected tubules called vesicular tubular clusters or VTCs. These clusters constitute a compartment at the ER-Golgi interface known as ERGIC (Endoplasmic Reticulum Golgi Intermediate Compartment). The ERGIC is a mobile membrane-bound cargo transport system that sorts proteins secreted from ER and delivers them to the Golgi.
With the help of motor proteins such...
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Transport Across the Golgi01:26

Transport Across the Golgi

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While it is unclear how molecules move between adjacent Golgi cisternae, it is apparent that the molecules move from cis- cisterna, the entry face, to the trans- cisterna, the exit face. Experiments initially suggested vesicles that bud from one cisterna and fuse with the next cisterna to transport proteins between the cisternae. This vesicular transport model describes the Golgi apparatus as a relatively static structure with a unique enzyme composition in each cisterna. Molecules are...
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Clathrin Coated Vesicles01:12

Clathrin Coated Vesicles

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Clathrin-coated vesicles use endocytosis to transport receptors and lysosomal hydrolases from the Golgi to the lysosome in the late secretory pathway. Clathrin-mediated endocytosis was the first described endocytic process, and Clathrin-coated vesicles remain one of the most well-studied transport vesicles. The molecular machinery that generates clathrin-coated vesicles comprises over 50 proteins that precisely coordinate vesicle formation. Cell surface receptors concentrated in indented sites...
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Video Experimental Relacionado

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In vivo and in vitro Studies of Adaptor-clathrin Interaction
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La carga regula la dinámica de las fosas recubiertas de clatrina.

Manojkumar A Puthenveedu1, Mark von Zastrow

  • 1Department of Psychiatry, University of California at San Francisco, 600 16th Street, San Francisco, CA 94158, USA.

Cell
|October 5, 2006
PubMed
Resumen

La endocitosis regulada de los receptores acoplados a proteínas G (GPCR) utiliza un subconjunto específico de fosas recubiertas de clatrina (CCP). Esta regulación específica de la carga de las CCP permite la especialización funcional en la vía endocítica.

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Área de la Ciencia:

  • Biología celular Biología celular.
  • Biología Molecular Biología Molecular
  • La bioquímica es la bioquímica.

Sus antecedentes:

  • Las fosas recubiertas de clatrina (CCP) son máquinas endocíticas clave.
  • Las CCP suelen verse como una población uniforme que maneja diversas cargas de membrana.
  • Los mecanismos específicos que rigen la segregación de carga y la dinámica de las CCP siguen siendo incompletamente entendidos.

Objetivo del estudio:

  • Para investigar si cargas específicas, como los receptores acoplados a proteínas G (GPCR), utilizan subconjuntos distintos de CCP para la endocitosis.
  • Para determinar si las ECC que contienen GPCR exhiben propiedades funcionales únicas.
  • Aclarar los mecanismos regulatorios que controlan la dinámica de las CCP basadas en el contenido de la carga.

Principales métodos:

  • Imágenes de células vivas para observar la dinámica de la CCP y el tráfico de carga.
  • Ensayos bioquímicos para analizar las interacciones proteína-proteína, incluidos los enlaces mediados por el dominio PDZ.
  • Estudios de perturbación para evaluar el impacto de la carga en el montaje y desmontaje de CCP.

Principales resultados:

  • La endocitosis regulada de los GPCR se produce preferentemente a través de un subconjunto especializado de CCP.
  • Las CCPs que contienen GPCR exhiben tiempos de residencia superficiales prolongados, regulados por las interacciones del citoesqueleto carga-actina.
  • Estos CCP especializados muestran un retraso en el reclutamiento de dinamina y pueden experimentar eventos abortivos, desacoplando el desmontaje de la capa de la internalización de la carga.

Conclusiones:

  • La carga endocítica, como los GPCR, puede segregarse en distintos subconjuntos de CCP.
  • La regulación dependiente de la carga de la dinámica CCP proporciona un mecanismo cinético para la especialización funcional.
  • Esta especialización optimiza la vía endocítica temprana y mitiga la competencia entre los diferentes tipos de carga.