A role for epsin N-terminal homology/AP180 N-terminal homology (ENTH/ANTH) domains in tubulin binding.
The epsin N-terminal homology (ENTH) domain is a protein module of approximately 150 amino acids found at the N terminus of a variety of proteins identified in yeast, plants, nematode, frog, and mammals. ENTH domains comprise multiple alpha-helices folded upon each other to form a compact globular structure that has been implicated in interactions with lipids and proteins. In characterizing this evolutionarily conserved domain, we isolated and identified tubulin as an ENTH domain-binding partner. The interaction, which is direct and has a dissociation constant of approximately 1 microm, was observed with ENTH domains of proteins present in various species. Tubulin is co-immunoprecipitated from rat brain extracts with the ENTH domain-containing proteins, epsins 1 and 2, and punctate epsin staining is observed along the microtubule cytoskeleton of dissociated cortical neurons. Consistent with a role in microtubule processes, the over-expression of epsin ENTH domain in PC12 cells stimulates neurite outgrowth. These data demonstrate an evolutionarily conserved property of ENTH domains to interact with tubulin and microtubules.
Hussain
Natasha K
NK
Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, Montreal, Quebec H3A 2B4, Canada.
Yamabhai
Montarop
M
Bhakar
Asha L
AL
Metzler
Martina
M
Ferguson
Stephen S G
SS
Hayden
Michael R
MR
McPherson
Peter S
PS
Kay
Brian K
BK
D033942:Adaptor Proteins, Vesicular Transport; D000818:Animals; D001665:Binding Sites; D001923:Brain Chemistry; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D017124:Conserved Sequence; D006801:Humans; D007163:Immunosorbent Techniques; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism; D034041:Monomeric Clathrin Assembly Proteins / Q000737:chemistry* / Q000378:metabolism; D016501:Neurites / Q000502:physiology; D009474:Neurons / Q000737:chemistry / Q000648:ultrastructure; D009479:Neuropeptides / Q000737:chemistry* / Q000378:metabolism; D017510:Protein Folding; D017433:Protein Structure, Secondary; D051381:Rats; D011993:Recombinant Fusion Proteins; D040681:Structural Homology, Protein; D014404:Tubulin / Q000032:analysis / Q000378:metabolism*; D033921:Vesicular Transport Proteins
D033942:Adaptor Proteins, Vesicular Transport; D002352:Carrier Proteins; D034041:Monomeric Clathrin Assembly Proteins; D009479:Neuropeptides; D011993:Recombinant Fusion Proteins; D014404:Tubulin; D033921:Vesicular Transport Proteins; C439023:clathrin assembly protein AP180; C114154:epsin