PMID- 10459011
OWN - NLM
STAT- MEDLINE
DCOM- 19990923
LR  - 20191023
IS  - 0021-9525 (Print)
IS  - 0021-9525 (Linking)
VI  - 146
IP  - 4
DP  - 1999 Aug 23
TI  - Phosphoinositide-AP-2 interactions required for targeting to plasma membrane
      clathrin-coated pits.
PG  - 755-64
AB  - The clathrin-associated AP-2 adaptor protein is a major
      polyphosphoinositide-binding protein in mammalian cells. A high affinity binding 
      site has previously been localized to the NH(2)-terminal region of the AP-2 alpha
      subunit (Gaidarov et al. 1996. J. Biol. Chem. 271:20922-20929). Here we used
      deletion and site- directed mutagenesis to determine that alpha residues 21-80
      comprise a discrete folding and inositide-binding domain. Further, positively
      charged residues located within this region are involved in binding, with a
      lysine triad at positions 55-57 particularly critical. Mutant peptides and
      protein in which these residues were changed to glutamine retained wild-type
      structural and functional characteristics by several criteria including circular 
      dichroism spectra, resistance to limited proteolysis, and clathrin binding
      activity. When expressed in intact cells, mutated alpha subunit showed defective 
      localization to clathrin-coated pits; at high expression levels, the appearance
      of endogenous AP-2 in coated pits was also blocked consistent with a
      dominant-negative phenotype. These results, together with recent work indicating 
      that phosphoinositides are also critical to ligand-dependent recruitment of
      arrestin-receptor complexes to coated pits (Gaidarov et al. 1999. EMBO (Eur. Mol.
      Biol. Organ.) J. 18:871-881), suggest that phosphoinositides play a critical and 
      general role in adaptor incorporation into plasma membrane clathrin-coated pits.
FAU - Gaidarov, I
AU  - Gaidarov I
AD  - Kimmel Cancer Institute and the Department of Microbiology and Immunology, Thomas
      Jefferson University, Philadelphia, Pennsylvania 19107, USA.
FAU - Keen, J H
AU  - Keen JH
LA  - eng
GR  - GM-49217/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Cell Biol
JT  - The Journal of cell biology
JID - 0375356
RN  - 0 (Adaptor Protein Complex alpha Subunits)
RN  - 0 (Adaptor Proteins, Vesicular Transport)
RN  - 0 (Clathrin)
RN  - 0 (Membrane Proteins)
RN  - 0 (Phosphatidylinositols)
RN  - 0RH81L854J (Glutamine)
RN  - K3Z4F929H6 (Lysine)
SB  - IM
MH  - Adaptor Protein Complex alpha Subunits
MH  - Adaptor Proteins, Vesicular Transport
MH  - Amino Acid Sequence
MH  - Amino Acid Substitution
MH  - Animals
MH  - Binding Sites
MH  - Biological Transport
MH  - Brain
MH  - Cattle
MH  - Cell Line
MH  - Cell Membrane/*metabolism
MH  - Circular Dichroism
MH  - Clathrin/*metabolism
MH  - Coated Pits, Cell-Membrane/*metabolism
MH  - Glutamine/genetics
MH  - Lysine/genetics/metabolism
MH  - Membrane Proteins/chemistry/genetics/*metabolism
MH  - Mice
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Phosphatidylinositols/*metabolism
MH  - Protein Structure, Secondary
MH  - Sequence Deletion
MH  - Transfection
PMC - PMC2156139
EDAT- 1999/08/25 00:00
MHDA- 1999/08/25 00:01
CRDT- 1999/08/25 00:00
PHST- 1999/08/25 00:00 [pubmed]
PHST- 1999/08/25 00:01 [medline]
PHST- 1999/08/25 00:00 [entrez]
AID - 10.1083/jcb.146.4.755 [doi]
PST - ppublish
SO  - J Cell Biol. 1999 Aug 23;146(4):755-64. doi: 10.1083/jcb.146.4.755.