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PDBsum entry 1k4z
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Membrane protein
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PDB id
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1k4z
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Contents |
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* Residue conservation analysis
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References listed in PDB file
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Key reference
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Title
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Crystal structure of the actin binding domain of the cyclase-Associated protein.
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Authors
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T.Dodatko,
A.A.Fedorov,
M.Grynberg,
Y.Patskovsky,
D.A.Rozwarski,
L.Jaroszewski,
E.Aronoff-Spencer,
E.Kondraskina,
T.Irving,
A.Godzik,
S.C.Almo.
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Ref.
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Biochemistry, 2004,
43,
10628-10641.
[DOI no: ]
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PubMed id
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Abstract
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Cyclase-associated protein (CAP or Srv2p) is a modular actin monomer binding
protein that directly regulates filament dynamics and has been implicated in a
number of complex developmental and morphological processes, including mRNA
localization and the establishment of cell polarity. The crystal structure of
the C-terminal dimerization and actin monomer binding domain (C-CAP) reveals a
highly unusual dimer, composed of monomers possessing six coils of right-handed
beta-helix flanked by antiparallel beta-strands. Domain swapping, involving the
last two strands of each monomer, results in the formation of an extended dimer
with an extensive interface. This structural and biochemical characterization
provides new insights into the organization and potential mechanistic properties
of the multiprotein assemblies that integrate dynamic actin processes into the
overall physiology of the cell. An unanticipated finding is that the unique
tertiary structure of the C-CAP monomer provides a structural model for a wide
range of molecules, including RP2 and cofactor C, proteins involved in X-linked
retinitis pigmentosa and tubulin maturation, respectively, as well as several
uncharacterized proteins that exhibit very diverse domain organizations. Thus,
the unusual right-handed beta-helical fold present in C-CAP appears to support a
wide range of biological functions.
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Secondary reference #1
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Title
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Two separate functions are encoded by the carboxyl-Terminal domains of the yeast cyclase-Associated protein and its mammalian homologs. Dimerization and actin binding.
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Authors
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A.Zelicof,
V.Protopopov,
D.David,
X.Y.Lin,
V.Lustgarten,
J.E.Gerst.
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Ref.
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J Biol Chem, 1996,
271,
18243-18252.
[DOI no: ]
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PubMed id
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Figure 4.
Fig. 4. The carboxyl terminus of both CAP and MCH1 binds
actin. A, full-length MCH1 and CAP bind actin. Lysates from  cap cells
(SKN50) expressing either Myc-tagged CAP or Myc-tagged MCH1 were
subjected to immunoprecipitation with anti-Myc antibody (9E10).
Specific protein interactions were blocked by the addition of
excess Myc peptide (50 µg). Immune complexes were resolved
on SDS-acylamide gels and transferred to nylon membranes.
Immunoblots were probed with an anti-actin monoclonal antibody
(20 µg/ml) (Boehringer Mannheim). B, the carboxyl-terminal
domain of CAP and MCH1 binds actin. Lysates from cap cells
expressing various Myc-tagged domains of CAP, or the
carboxyl-terminal of MCH1, were subjected to
immunoprecipitation. Immunoblots were incubated with an
anti-actin monoclonal antibody (20 µg/ml). Antigen
detection was performed by ECL chemiluminescent assay.
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Figure 5.
Fig. 5. The actin binding and dimerization functions of CAP
are mediated by separate domains. A, the last 27 amino acids of
CAP are essential for actin binding. cap yeast
cells (SKN32) bearing plasmids expressing HA-CAP (YCpADH-HACAP)
and Myc-CAP1-498 (pADH-mycCAP 11) were
grown to log phase, prior to harvesting, and lysate preparation.
B, CAP1-498 forms homodimers. cap yeast
cells (SKN32) bearing plasmids expressing HA-CAP1-498
(pTADH-HACAP 11) and
Myc-CAP1-498 (pADH-mycCAP 11) were
grown to log phase, prior to harvesting and lysate preparation.
Immunoprecipitations were carried out, as described under
``Experimental Procedures,'' using either anti-HA ascites
(12CA5) or anti-Myc antisera (9E10). Specific protein-protein
interactions were blocked by the addition of either excess HA
(30 µg) or Myc peptide (30 µg) to
immunoprecipitation reactions (+peptide). Detection of tagged
CAP molecules or actin was accomplished using the appropriate
monoclonal antibodies (anti-actin (6 µg/ml), anti-HA
ascites fluid (1:5000), and anti-Myc (1:5000)). Lanes marked TCL
contained 75 µg of total cell lysate. Antigen detection
was performed by ECL chemiluminescent assay.
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The above figures are
reproduced from the cited reference
with permission from the ASBMB
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