title: Comparative analysis of adaptor-mediated clathrin assembly reveals general principles for adaptor clustering.
abstract: Clathrin-mediated endocytosis (CME) manages the sorting and uptake of the bulk of membrane proteins (or cargo) from the plasma membrane. CME is initiated by the formation of clathrin-coated pits (CCPs), in which adaptors nucleate clathrin assembly. Clathrin adaptors display diversity in both the type and number of evolutionarily conserved clathrin-binding boxes. How this diversity relates to the process of adaptor clustering as clathrin assembles around a growing pit remains unclear. Using real-time, fluorescence microscopy-based assays, we compare the formation kinetics and distribution of clathrin assemblies on membranes that display five unique clathrin adaptors. Correlations between equilibrium and kinetic parameters of clathrin assembly to the eventual adaptor distribution indicate that adaptor clustering is determined not by the amount of clathrin recruited or the degree of clathrin clustered but instead by the rate of clathrin assembly. Together our results emphasize the need to analyze kinetics of protein interactions to better understand mechanisms that regulate CME.
authors: Indian Institute of Science Education and Research, Pune, Maharashtra 411 008, India pucadyil@iiserpune.ac.in.
authors: Thomas J
authors: TJ
authors: Pucadyil
authors: Indian Institute of Science Education and Research, Pune, Maharashtra 411 008, India.
authors: Sachin S
authors: SS
authors: Holkar
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism* / Q000502:physiology; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002966:Clathrin / Q000378:metabolism; D022163:Clathrin-Coated Vesicles / Q000378:metabolism*; D003034:Coated Pits, Cell-Membrane / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology; D006801:Humans; D007700:Kinetics; D008565:Membrane Proteins / Q000378:metabolism; D061848:Optical Imaging / Q000379:methods; D011485:Protein Binding; D021381:Protein Transport / Q000502:physiology
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D002966:Clathrin; D008565:Membrane Proteins