The two-step cargo recognition mechanism of heterotrimeric kinesin.

Jiang X, Ogawa T, Yonezawa K, Shimizu N, Ichinose S, Uchihashi T, Nagaike W, Moriya T, Adachi N, Kawasaki M, Dohmae N, Senda T, Hirokawa N, EMBO Rep :e56864 (2023) Europe PMC

SASDMW5 – Kinesin superfamily protein 3 A/B and kinesin associated protein 3 (KAP) with APC ARM

Adenomatous polyposis coli protein (N-terminal ARM domain)
Kinesin-like protein KIF3A
Kinesin-like protein KIF3B
Kinesin-associated protein 3
MWI(0) 264 kDa
MWexpected 216 kDa
VPorod 763 nm3
log I(s) 1.78×10-1 1.78×10-2 1.78×10-3 1.78×10-4
Adenomatous polyposis coli protein (N-terminal ARM domain) Kinesin-like protein KIF3A Kinesin-like protein KIF3B Kinesin-associated protein 3 small angle scattering data  s, nm-1
ln I(s)
Adenomatous polyposis coli protein (N-terminal ARM domain) Kinesin-like protein KIF3A Kinesin-like protein KIF3B Kinesin-associated protein 3 Guinier plot ln 1.78×10-1 Rg: 5.4 nm 0 (5.4 nm)-2 s2
(sRg)2I(s)/I(0)
Adenomatous polyposis coli protein (N-terminal ARM domain) Kinesin-like protein KIF3A Kinesin-like protein KIF3B Kinesin-associated protein 3 Kratky plot 1.104 0 √3 sRg
p(r)
Adenomatous polyposis coli protein (N-terminal ARM domain) Kinesin-like protein KIF3A Kinesin-like protein KIF3B Kinesin-associated protein 3 pair distance distribution function Rg: 5.4 nm 0 Dmax: 19.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Adenomatous polyposis coli protein (N-terminal ARM domain) Kinesin-like protein KIF3A Kinesin-like protein KIF3B Kinesin-associated protein 3 DAMMIF model
Synchrotron SAXS data from solutions of Kinesin superfamily protein 3 A/B and Kinesin associated protein 3 with the N-terminal ARM domain of Adenomatous polyposis coli protein in 25 mM TrisHCl, 200 mM NaCl, 5% glycerol, 1mM DTT pH 8 were collected on the beam line BL-10C at the Phton Factory (PF; Tsukuba, Ibaraki, Japan) using a Pilatus3 2M detector at a sample-detector distance of 2.08 m and at a wavelength of λ = 0.1 nm (I(s) vs s, where s = 4πsinθ/λ and 2θ is the scattering angle). 540 successive 20 second frames were collected at 20°C using size-exclusion chromatography SAXS. Sample was loaded to Superdex 200 increase 10/300 (GE healthcare). Flow rate was set at 0.05 mL/min. The data were normalized to the intensity of the incident beam and circularly averaged; the scattering of the solvent-blank was subtracted. The extrapolated scattering profile at a concentration of zero was calculated with the data near the peak top above half (frames 206–244) by Serial Analyzer. In order to estimate the molecular mass from I(0), PSV (0.734 cm3/g) and the scattering mass contrast (2.763×10^10 cm-2) of this protein were calculated by using MULCh (http://smb-research.smb.usyd.edu.au/NCVWeb/input.jsp).

Adenomatous polyposis coli protein (N-terminal ARM domain) (APC-ARM)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   75.3 kDa
 
UniProt   Q61315 (338-1010)
Sequence   FASTA
 
Kinesin-like protein KIF3A (KIF 3 A H)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   27.9 kDa
 
UniProt   P28741 (481-701)
Sequence   FASTA
 
Kinesin-like protein KIF3B (KIF 3B)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   31.9 kDa
 
UniProt   Q61771 (475-747)
Sequence   FASTA
 
Kinesin-associated protein 3 (KAP 3)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   81.2 kDa
 
UniProt   P70188 (1-692)
Sequence   FASTA