Solution Structure of Eps15's Third EH Domain Reveals Coincident Phe−Trp and Asn−Pro−Phe Binding Sites†,‡

Biochemistry ◽  
2000 ◽  
Vol 39 (15) ◽  
pp. 4309-4319 ◽  
Author(s):  
Jennifer L. Enmon ◽  
Tonny de Beer ◽  
Michael Overduin
Keyword(s):  
Binding Sites ◽  
Solution Structure ◽  
Eh Domain

scholarly journals Structure-Based Functional Analysis of a Hormone Belonging to an Ecdysozoan Peptide Superfamily: Revelation of a Common Molecular Architecture and Residues Possibly for Receptor Interaction

2021 ◽  
Vol 22 (20) ◽  
pp. 11142
Author(s):  
Yun-Ru Chen ◽  
Nai-Wan Hsiao ◽  
Yi-Zong Lee ◽  
Shiau-Shan Huang ◽  
Chih-Chun Chang ◽  
...  
Keyword(s):  
Atpase Activity ◽  
Receptor Binding ◽  
Binding Sites ◽  
Disulfide Bonds ◽  
Solution Structure ◽  
Receptor Interaction ◽  
Resonance Spectroscopy ◽  
Molecular Architecture ◽  
Major Factors

A neuropeptide (Sco-CHH-L), belonging to the crustacean hyperglycemic hormone (CHH) superfamily and preferentially expressed in the pericardial organs (POs) of the mud crab Scylla olivacea, was functionally and structurally studied. Its expression levels were significantly higher than the alternative splice form (Sco-CHH) in the POs, and increased significantly after the animals were subjected to a hypo-osmotic stress. Sco-CHH-L, but not Sco-CHH, significantly stimulated in vitro the Na+, K+-ATPase activity in the posterior (6th) gills. Furthermore, the solution structure of Sco-CHH-L was resolved using nuclear magnetic resonance spectroscopy, revealing that it has an N-terminal tail, three α-helices (α2, Gly9−Asn28; α3, His34−Gly38; and α5, Glu62−Arg72), and a π-helix (π4, Cys43−Tyr54), and is structurally constrained by a pattern of disulfide bonds (Cys7–Cys43, Cys23–Cys39, and Cys26–Cys52), which is characteristic of the CHH superfamily-peptides. Sco-CHH-L is topologically most similar to the molt-inhibiting hormone from the Kuruma prawn Marsupenaeus japonicus with a backbone root-mean-square-deviation of 3.12 Å. Ten residues of Sco-CHH-L were chosen for alanine-substitution, and the resulting mutants were functionally tested using the gill Na+, K+-ATPase activity assay, showing that the functionally important residues (I2, F3, E45, D69, I71, and G73) are located at either end of the sequence, which are sterically close to each other and presumably constitute the receptor binding sites. Sco-CHH-L was compared with other members of the superfamily, revealing a folding pattern, which is suggested to be common for the crustacean members of the superfamily, with the properties of the residues constituting the presumed receptor binding sites being the major factors dictating the ligand–receptor binding specificity.

scholarly journals Solution Structure of CCL19 and Identification of Overlapping CCR7 and PSGL-1 Binding Sites

Biochemistry ◽  
2015 ◽  
Vol 54 (27) ◽  
pp. 4163-4166 ◽  
Author(s):  
Christopher T. Veldkamp ◽  
Eva Kiermaier ◽  
Skylar J. Gabel-Eissens ◽  
Miranda L. Gillitzer ◽  
David R. Lippner ◽  
...  
Keyword(s):  
Binding Sites ◽  
Solution Structure

scholarly journals The clathrin adaptor Dab2 recruits EH domain scaffold proteins to regulate integrin β1 endocytosis

2012 ◽  
Vol 23 (15) ◽  
pp. 2905-2916 ◽  
Author(s):  
Anjali Teckchandani ◽  
Erin E. Mulkearns ◽  
Timothy W. Randolph ◽  
Natalie Toida ◽  
Jonathan A. Cooper
Keyword(s):  
Transferrin Receptor ◽  
Binding Sites ◽  
Adaptor Proteins ◽  
Scaffold Proteins ◽  
Receptor Internalization ◽  
Accessory Proteins ◽  
Integrin Β1 ◽  
Eh Domain ◽  
Pit Nucleation ◽  
Clathrin Adaptor

Endocytic adaptor proteins facilitate cargo recruitment and clathrin-coated pit nucleation. The prototypical clathrin adaptor AP2 mediates cargo recruitment, maturation, and scission of the pit by binding cargo, clathrin, and accessory proteins, including the Eps-homology (EH) domain proteins Eps15 and intersectin. However, clathrin-mediated endocytosis of some cargoes proceeds efficiently in AP2-depleted cells. We found that Dab2, another endocytic adaptor, also binds to Eps15 and intersectin. Depletion of EH domain proteins altered the number and size of clathrin structures and impaired the endocytosis of the Dab2- and AP2-dependent cargoes, integrin β1 and transferrin receptor, respectively. To test the importance of Dab2 binding to EH domain proteins for endocytosis, we mutated the EH domain–binding sites. This mutant localized to clathrin structures with integrin β1, AP2, and reduced amounts of Eps15. Of interest, although integrin β1 endocytosis was impaired, transferrin receptor internalization was unaffected. Surprisingly, whereas clathrin structures contain both Dab2 and AP2, integrin β1 and transferrin localize in separate pits. These data suggest that Dab2-mediated recruitment of EH domain proteins selectively drives the internalization of the Dab2 cargo, integrin β1. We propose that adaptors may need to be bound to their cargo to regulate EH domain proteins and internalize efficiently.

scholarly journals Solution Structure of a Repeated Unit of the ABA-1 Nematode Polyprotein Allergen of Ascaris Reveals a Novel Fold and Two Discrete Lipid-Binding Sites

2011 ◽  
Vol 5 (4) ◽  
pp. e1040 ◽  
Author(s):  
Nicola A. G. Meenan ◽  
Graeme Ball ◽  
Krystyna Bromek ◽  
Dušan Uhrín ◽  
Alan Cooper ◽  
...  
Keyword(s):  
Binding Sites ◽  
Solution Structure ◽  
Lipid Binding ◽  
Repeated Unit

scholarly journals The solution structure of the transducin-α-uncoordinated 119 protein complex suggests occlusion of the Gβ1γ1-binding sites

FEBS Journal ◽  
2014 ◽  
Vol 282 (3) ◽  
pp. 550-561 ◽  
Author(s):  
Pallavi Cheguru ◽  
Anurima Majumder ◽  
Ravi Yadav ◽  
Kota N. Gopalakrishna ◽  
Lokesh Gakhar ◽  
...  
Keyword(s):  
Protein Complex ◽  
Binding Sites ◽  
Solution Structure

Solution Structure of the Reps1 EH Domain and Characterization of Its Binding to NPF Target Sequences†,‡

Biochemistry ◽  
2001 ◽  
Vol 40 (23) ◽  
pp. 6776-6785 ◽  
Author(s):  
Soyoun Kim ◽  
Donald N. Cullis ◽  
Larry A. Feig ◽  
James D. Baleja
Keyword(s):  
Solution Structure ◽  
Eh Domain ◽  
Target Sequences
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