Mechanochemical properties of brain clathrin: interactions with actin and alpha-actinin and polymerization into basketlike structures or filaments.

Schook, William; Puszkin, Saul; Bloom, William; Ores, Christine; Kochwa, Shaul

doi:10.1073/pnas.76.1.116

Cited by 97 publications

(82 citation statements)

References 15 publications

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“…We have demonstrated also that baskets are not formed in the presence of high concentrations of divalent cations and salts [5]. Here we have combined both observations to monitor clathrin polymerization upon addition of these salts.…”

Section: Discussionmentioning

confidence: 95%

“…The basic procedure used for adsorption of proteins to these particles was described elsewhere [5]. Clathrin-coated polystyrene particles were suspended in a 0.5-ml solution of 0.5 mM chlorpromazine, pH 6.5, and washed twice to remove unbound drug before the binding of a second protein was at tempted.…”

Section: Lytron Polystyrene Particle Bindingmentioning

confidence: 99%

“…We reported earlier that purified clathrin is capable of reversible assembly in vitro simply by lowering its pH in solution from 7.5 to 6.5 [ 3 ] . At pH 6.5 clathrin polymerizes into basket-like or cage-like structures, whose lattices are similar in size and shape to those on coated vesicles observed by electron microscopy [5]. The increased turbidity that resulted when baskets formed in solution at pH 6.5 was attributed to the light-scattering ability of the cage structures [6].…”

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confidence: 99%

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Brain Clathrin: Studies of Its Ultrastructural Assemblies

et al. 1982

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Clathrin p u r i h d to a high dzgrec of hoinogzncity showed thz presencz of accompanying polypzptidcs of lowsr molccular wcights and asscmblitd into baskcls or c a g a when thc pH of its solution was adjusted from pH 7.5 to 6.5. Brief chyinotrypsin treatment of this clathrin preparation at pH 6.5 cleaved clathrin-associated proteins rendzring clathrin unablc to rcform baskets. Hydrolysis of clathrin-associated proteins did not afYect clathrin's capacity to polymerize into open latticzs or to bind actin and a-actinin from solution. W1ii.n open latticcs formzd, th; turbidity of the solution rose to lzvels that wcr2 higha than t1ios;'produced when cages or baskets wirc: form2d by native clathrin. In both native or enzyme-treated clathrin, turbidity increase was inhibited by salts. The addition of ccrtain cytoskzlctal-disrupting agents, such a s chlorpromazinit and imipraminz, incrzascd clathrin's turbidit y but did not r.:sult in formation of baskets. Colchicinz and cytochalasin B did not affect turbidity or t h s asszmbly o f cagzs. Thz addition of incrcasing ainounls of vinblastin,: rcsulted in the prxipitation of larger amounts of clatlirin which, after solubilization, r=taincd its ability to polymierize into baskets. I t sJems thal clathrin (exists as a complex with other protein moleculcs, clatlirin-associatzd protdins, that inodulatz tht: cxtznt of polyincrization and asscmbly of clathrin into charactcristic structurzs.

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Section: Discussionmentioning

confidence: 95%

Section: Lytron Polystyrene Particle Bindingmentioning

confidence: 99%

mentioning

confidence: 99%

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Brain Clathrin: Studies of Its Ultrastructural Assemblies

et al. 1982

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“…It not only binds with high affinity to actin microfilaments and other proteins (56) but also associates with specialized membranes (50,51). Clathrin can self-assemble (31), and it may exist in more than one structural form (17,56). These properties suggest that clathrin could be a multifunctional component ofthe carrier structure .…”

Section: Identification Of the Proteins Comprising The Carrier Structmentioning

confidence: 99%

Axonal transport of the cytoplasmic matrix.

Lasek¹,

Garner²,

Brady³

1984

The Journal of Cell Biology

284

155

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The cytoplasmic matrix is often highly specialized, making it possible to clearly relate particularaspects ofthe cytoplasmic matrix to the specialized functions of cells . For example, in striated muscle cells the contractile components of the cytoplasmic matrix dominate the cell structurally and functionally. Neurons are another example of cells in which specializations of structure and function can be clearly related to particular aspects of the cytoplasmic matrix (36, 37). The primary function of neurons is to convey information from one location in the organism to another. Pathways for information transfer in the nervous system are provided by specialized neuronal extensions, the axons and the dendrites . Axons, in particular, are specialized to convey information over very long distances, meters in some cases. Accordingly, in the axon the cytoplasmic matrix is specialized to generate and support the extremely elongate shape ofthe axon during development, regeneration, and maturity.To generate and maintain the great volume of cytoplasm within the axon, neurons must produce tremendous amounts of protein (32). Essentially all axonal proteins are synthesized in the neuron cell body and then conveyed into the axon by axonal transport, which provides a lifeline for the axon and its terminus (25,36). Axonal transport is a process that is initiated when the axon first develops and that continues throughout the life of the neuron. To meet the needs of large animals, which require long axons, axonal transport has become one of the most highly developed mechanisms for the intracellular transport of materials in metazoan cells .

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“…Purified bovine brain CCV [18], synaptic vesicles [19] and synaptic plasma membranes [19] were prepared as described. Enzyme profiles of the membrane fractions were reported elsewhere [20].…”

Section: Preparation Of Membrane Fractionsmentioning

confidence: 99%

Novel proteins mediate an interaction between clathrin‐coated vesicles and polymerizing actin filaments

Kohtz

Hanson

Puszkin

1990

European Journal of Biochemistry

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A monoclonal antibody, A-7C11, was generated which reacts with two polypeptides of 40 kDa and 80 kDa associated with the coat proteins of purified brain clathirn-coated vesicles. The 40-kDa antigen was purified and found to display actin-binding properties. Negative-staining electron microscopy showed that one of the antigens reactive with A-7C11 appears to mediate the association of isolated clathrin-coated vesicles with assembling actin filaments in vitro. Immunofluorescence microscopy of cultured fibroblasts with A-7C11 revealed the antigens aligned with both actin filaments and as punctate structures near the plasma membrane. The data suggest that the interaction between clathrin-coated vesicles and the actin cytoskeleton is mediated by antigens identified by monoclonal antibody A-7Cll.Clathrin-coated vesicles (CCV) associated with the plasma membrane participate in the clustering and internalization of receptor-ligand complexes [l, 21. Those associated with the Golgi cisternae mediate the processing [3] and secretion [4] of certain polypeptides, while those associated with synaptic terminals [2, 51 and the secretory regions of cells [6] may be involved in membrane recovery. Clathrin-coated vesicles have been isolated from a variety of tissues, and some of the major protein components of the vesicles' protein coat have been described (reviewed in [7]). On the other hand, a coherent understanding of how CCV participate in selecting their cargo, navigate the subcellular milieu, identify and interact with their target organelles has not emerged.In previous reports we showed that clathrin coat proteins displayed binding affinity for structural, anchoring and regulatory molecules of the cytoskeleton, namely actin, a-actinin and calmodulin [8, 91. More recently, the binding sites on clathrin for a-actinin and calmodulin were also defined [lo]. Immunofluorescence and ultrastructural studies by us and others suggested that a correlation exists in the distribution of clathrin-coated vesicles with actin microfilaments [11 -141. Moreover, a neuronal protein of 185 kDa (NP185), found associated with bovine brain clathrin-coated vesicles, displayed binding affinity for tubulin which was regulated by the casein kinase I1 phosphorylation of tubulin [15, 161. Thus, the identification of novel molecules in CCV which could mediate and regulate clathrin-coated vesicle functions related to the cytoskeleton could yield valuable insights into the biochemical basis of selective molecules and membrane transport.The generation of monoclonal antibodies (mAb) provided sensitive probes to identify and characterize individual components present in different types of vesicles [17] ual components of the mixture were then separated by the generation and cloning of hybridoma cells. The mAb A-7C11 was generated in this manner, using the dissociated coat proteins of bovine brain CCV as an immunogen. Among them, two previously unobserved components of 40 kDa and 80 kDa present in bovine brain CCV were found to be the cross-reactive antignes of...

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Mechanochemical properties of brain clathrin: interactions with actin and alpha-actinin and polymerization into basketlike structures or filaments.

Cited by 97 publications

References 15 publications

Brain Clathrin: Studies of Its Ultrastructural Assemblies

Brain Clathrin: Studies of Its Ultrastructural Assemblies

Axonal transport of the cytoplasmic matrix.

Novel proteins mediate an interaction between clathrin‐coated vesicles and polymerizing actin filaments

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