2WOZ

The novel beta-propeller of the BTB-Kelch protein Krp1 provides the binding site for Lasp-1 that is necessary for pseudopodia extension

PDB DOI: https://doi.org/10.2210/pdb2WOZ/pdb

Classification: PROTEIN BINDING
Organism(s): Rattus norvegicus
Expression System: Escherichia coli BL21(DE3)
Mutation(s): No

Deposited: 2009-07-31 Released: 2009-09-01
Deposition Author(s): Gray, C.H., McGarry, L.C., Spence, H.J., Riboldi-Tunnicliffe, A., Ozanne, B.W.

Experimental Data Snapshot

Method: X-RAY DIFFRACTION
Resolution: 2.00 Å
R-Value Free: 0.279
R-Value Work: 0.237

wwPDB Validation 3D Report Full Report

This is version 1.3 of the entry. See complete history.

Literature

Novel beta-propeller of the BTB-Kelch protein Krp1 provides a binding site for Lasp-1 that is necessary for pseudopodial extension.

Gray, C.H., McGarry, L.C., Spence, H.J., Riboldi-Tunnicliffe, A., Ozanne, B.W.

(2009) J Biol Chem 284: 30498-30507

PubMed: 19726686 Search on PubMedSearch on PubMed Central
DOI: https://doi.org/10.1074/jbc.M109.023259
Primary Citation of Related Structures:
2WOZ

PubMed Abstract:
Kelch-related protein 1 (Krp1) is up-regulated in oncogene-transformed fibroblasts. The Kelch repeats interact directly with the actin-binding protein Lasp-1 in membrane ruffles at the tips of pseudopodia, where both proteins are necessary for pseudopodial elongation. Herein, we investigate the molecular basis for this interaction. Probing an array of overlapping decapeptides of Rattus norvegicus (Rat) Krp1 with recombinant Lasp-1 revealed two binding sites; one ((317)YDPMENECYLT(327)) precedes the first of five Kelch repeats, and the other ((563)TEVNDIWKYEDD(574)) is in the last of the five Kelch repeats. Mutational analysis established that both binding sites are necessary for Krp1-Lasp-1 interaction in vitro and function in vivo. The crystal structure of the C-terminal domain of rat Krp1 (amino acids 289-606) reveals that both binding sites are brought into close proximity by the formation of a novel six-bladed beta-propeller, where the first blade is not formed by a Kelch repeat.

Organizational Affiliation

Invasion and Metastasis Laboratory, Beatson Institute for Cancer Research, Garscube Estate, Glasgow G61 1BD, Scotland, United Kingdom. c.gray@beatson.gla.ac.uk

Macromolecule Content

Total Structure Weight: 35.42 kDa
Atom Count: 2,556
Modelled Residue Count: 307
Deposited Residue Count: 318
Unique protein chains: 1

Macromolecules

Find similar proteins by:

(by identity cutoff) | 3D Structure

Entity ID: 1
Molecule	Chains	Sequence Length	Organism	Details	Image
KELCH REPEAT AND BTB DOMAIN-CONTAINING PROTEIN 10	A	318	Rattus norvegicus	Mutation(s): 0
UniProt
Find proteins for Q9ER30 (Rattus norvegicus) Explore Q9ER30 Go to UniProtKB: Q9ER30
Entity Groups
Sequence Clusters	30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt Group	Q9ER30
Sequence Annotations Expand
Reference Sequence

Experimental Data & Validation

Experimental Data

Method: X-RAY DIFFRACTION
Resolution: 2.00 Å
R-Value Free: 0.279
R-Value Work: 0.237
Space Group: P 2₁ 2₁ 2

Unit Cell:

Length ( Å )	Angle ( ˚ )
a = 66.11	α = 90
b = 98.53	β = 90
c = 46.97	γ = 90

Software Package:

Software Name	Purpose
REFMAC	refinement
MOSFLM	data reduction
SCALA	data scaling
SHARP	phasing

Structure Validation

View Full Validation Report

Entry History

Deposition Data

Released Date: 2009-09-01

Deposition Author(s):

Gray, C.H.

McGarry, L.C.

Spence, H.J.

Riboldi-Tunnicliffe, A.

Ozanne, B.W.

Revision History (Full details and data files)

Version 1.0: 2009-09-01
Type: Initial release
Version 1.1: 2011-07-07
Changes: Version format compliance
Version 1.2: 2011-07-13
Changes: Version format compliance
Version 1.3: 2018-06-13
Changes: Data collection, Database references, Source and taxonomy