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.
Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, Montreal, Quebec H3A 2B4, Canada.
Natasha K
NK
Hussain
Montarop
M
Yamabhai
Asha L
AL
Bhakar
Martina
M
Metzler
Stephen S G
SS
Ferguson
Michael R
MR
Hayden
Peter S
PS
McPherson
Brian K
BK
Kay
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