CALM regulates clathrin-coated vesicle size and maturation by directly sensing and driving membrane curvature.
CALM regulates clathrin-coated vesicle size and maturation by directly sensing and driving membrane curvature.
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DOI：
10.1016/j.devcel.2015.03.002
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发表时间：
2015-04-20
影响因子：
11.8
通讯作者：
Owen, David J.
中科院分区：
文献类型：
--
作者：
Miller, Sharon E.;Mathiasen, Signe;Bright, Nicholas A.;Pierre, Fabienne;Kelly, Bernard T.;Kladt, Nikolay;Schauss, Astrid;Merrifield, Christien J.;Stamou, Dimitrios;Hoening, Stefan;Owen, David J.
内吞网格蛋白包被囊泡（CCV）的大小是非常均匀的，这表明它是优化的，以实现适当水平的货物和脂质内化。哺乳动物内吞CCV中三种最丰富的蛋白质是网格蛋白和两种货物选择网格蛋白衔接子CALM和AP 2。在这里，我们表明，CALM的耗尽导致“开放”网格蛋白包被的凹坑（CCP）的比例大幅增加，以“颈缩”/“封闭”CCV和CCP/CCV直径的两倍，而AP 2耗尽具有相反的效果。然而，耗尽任一接头，显着抑制转铁蛋白和表皮生长因子的内吞作用。CALM耗竭的表型效应可以通过野生型CALM的再表达来挽救，但不能用缺乏功能性N-末端、膜插入、曲率传感/驱动两亲性螺旋的CALM来挽救，所述两亲性螺旋的存在和性质已被证明。因此，CALM是控制CCV大小和成熟的主要因素，因此决定了内吞货物摄取的速率。CALM损失增加了早期内吞网格蛋白包被结构的大小和频率CALM的耗尽减慢了内吞网格蛋白包被的小凹成熟和内吞速率CALM具有N-末端、膜曲率感应/驱动两亲性螺旋网格蛋白包被的小凹成熟由CALM的N-末端两亲性螺旋米勒等人调节。证明了丰富的内吞蛋白CALM的耗尽增加了早期内吞网格蛋白包被结构（CCS）的直径和百分比，同时延迟CCS成熟并降低内吞速率。CALM影响这些CCS特性的能力取决于插入膜的两亲性螺旋，该螺旋感测并促进膜曲率。
The size of endocytic clathrin-coated vesicles (CCVs) is remarkably uniform, suggesting that it is optimized to achieve the appropriate levels of cargo and lipid internalization. The three most abundant proteins in mammalian endocytic CCVs are clathrin and the two cargo-selecting, clathrin adaptors, CALM and AP2. Here we demonstrate that depletion of CALM causes a substantial increase in the ratio of “open” clathrin-coated pits (CCPs) to “necked”/“closed” CCVs and a doubling of CCP/CCV diameter, whereas AP2 depletion has opposite effects. Depletion of either adaptor, however, significantly inhibits endocytosis of transferrin and epidermal growth factor. The phenotypic effects of CALM depletion can be rescued by re-expression of wild-type CALM, but not with CALM that lacks a functional N-terminal, membrane-inserting, curvature-sensing/driving amphipathic helix, the existence and properties of which are demonstrated. CALM is thus a major factor in controlling CCV size and maturation and hence in determining the rates of endocytic cargo uptake. CALM loss increases size and frequency of early endocytic clathrin-coated structures Depletion of CALM slows endocytic clathrin-coated pit maturation and endocytic rate CALM possesses an N-terminal, membrane-curvature-sensing/driving amphipathic helix Clathrin-coated pit maturation is regulated by CALM’s N-terminal amphipathic helix Miller et al. demonstrate that depletion of the abundant endocytic protein CALM increases the diameter and percentage of early endocytic clathrin-coated structures (CCSs) while delaying CCS maturation and reducing endocytic rates. CALM’s ability to influence these CCS properties depends on a membrane-inserting amphipathic helix, which senses and promotes membrane curvature.
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DOI：
10.1083/jcb.84.3.560
发表时间：
1980-03
期刊：
The Journal of cell biology
影响因子：
--
作者：
Heuser J
通讯作者：
Heuser J
影响因子：
64.5
作者：
Boucrot E;Pick A;Çamdere G;Liska N;Evergren E;McMahon HT;Kozlov MM
通讯作者：
Kozlov MM
影响因子：
30.8
作者：
Harold, Denise;Abraham, Richard;Hollingworth, Paul;Sims, Rebecca;Gerrish, Amy;Hamshere, Marian L.;Pahwa, Jaspreet Singh;Moskvina, Valentina;Dowzell, Kimberley;Williams, Amy;Jones, Nicola;Thomas, Charlene;Stretton, Alexandra;Morgan, Angharad R.;Lovestone, Simon;Powell, John;Proitsi, Petroula;Lupton, Michelle K.;Brayne, Carol;Rubinsztein, David C.;Gill, Michael;Lawlor, Brian;Lynch, Aoibhinn;Morgan, Kevin;Brown, Kristelle S.;Passmore, Peter A.;Craig, David;McGuinness, Bernadette;Todd, Stephen;Holmes, Clive;Mann, David;Smith, A. David;Love, Seth;Kehoe, Patrick G.;Hardy, John;Mead, Simon;Fox, Nick;Rossor, Martin;Collinge, John;Maier, Wolfgang;Jessen, Frank;Schuermann, Britta;van den Bussche, Hendrik;Heuser, Isabella;Kornhuber, Johannes;Wiltfang, Jens;Dichgans, Martin;Froelich, Lutz;Hampel, Harald;Huell, Michael;Rujescu, Dan;Goate, Alison M.;Kauwe, John S. K.;Cruchaga, Carlos;Nowotny, Petra;Morris, John C.;Mayo, Kevin;Sleegers, Kristel;Bettens, Karolien;Engelborghs, Sebastiaan;De Deyn, Peter P.;Van Broeckhoven, Christine;Livingston, Gill;Bass, Nicholas J.;Gurling, Hugh;McQuillin, Andrew;Gwilliam, Rhian;Deloukas, Panagiotis;Al-Chalabi, Ammar;Shaw, Christopher E.;Tsolaki, Magda;Singleton, Andrew B.;Guerreiro, Rita;Muehleisen, Thomas W.;Noethen, Markus M.;Moebus, Susanne;Joeckel, Karl-Heinz;Klopp, Norman;Wichmann, H-Erich;Carrasquillo, Minerva M.;Pankratz, V. Shane;Younkin, Steven G.;Holmans, Peter A.;O'Donovan, Michael;Owen, Michael J.;Williams, Julie
通讯作者：
Williams, Julie
影响因子：
11.4
作者：
Bhatia, Vikram K.;Madsen, Kenneth L.;Stamou, Dimitrios
通讯作者：
Stamou, Dimitrios
影响因子：
64.8
作者：
Ford, MGJ;Mills, IG;McMahon, HT
通讯作者：
McMahon, HT