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

SASDMV5 – Kinesin superfamily protein 3 A/B and kinesin associated protein 3 (KAP)

Kinesin-like protein KIF3B
Kinesin-associated protein 3
Kinesin-like protein KIF3A
MWI(0) 227 kDa
MWexpected 141 kDa
VPorod 557 nm3
log I(s) 1.54×10-1 1.54×10-2 1.54×10-3 1.54×10-4
Kinesin-like protein KIF3B Kinesin-associated protein 3 Kinesin-like protein KIF3A small angle scattering data  s, nm-1
ln I(s)
Kinesin-like protein KIF3B Kinesin-associated protein 3 Kinesin-like protein KIF3A Guinier plot ln 1.55×10-1 Rg: 5.8 nm 0 (5.8 nm)-2 s2
(sRg)2I(s)/I(0)
Kinesin-like protein KIF3B Kinesin-associated protein 3 Kinesin-like protein KIF3A Kratky plot 1.104 0 √3 sRg
p(r)
Kinesin-like protein KIF3B Kinesin-associated protein 3 Kinesin-like protein KIF3A pair distance distribution function Rg: 6.3 nm 0 Dmax: 27 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Kinesin-like protein KIF3B Kinesin-associated protein 3 Kinesin-like protein KIF3A DAMMIF model
Synchrotron SAXS data from solutions of Kinesin superfamily protein 3 A/B and Kinesin associated protein 3 in 20 mM TrisHCl, 200 mM NaCl, 5% Glycerol, 1mM DTT pH 8 were collected on the beam line BL-15A2 at the Phton Factory (PF; Tsukuba, Ibaraki, Japan) using a Pilatus3 2M detector at a sample-detector distance of 2.65 m and at a wavelength of λ = 0.1 nm (I(s) vs s, where s = 4πsinθ/λ and 2θ is the scattering angle). 815 successive 5 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 0.1 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 in the descent side of the peak (frames 325–344) by Serial Analyzer. In order to estimate the molecular mass from I(0), PSV (0.739 cm3/g) and the scattering mass contrast (2.698×10^10 cm-2) of this protein were calculated by using MULCh (http://smb-research.smb.usyd.edu.au/NCVWeb/input.jsp).

Kinesin-like protein KIF3B (KIF 3 B)
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   81.1 kDa
 
UniProt   P70188 (1-693)
Sequence   FASTA
 
Kinesin-like protein KIF3A (KIF 3 A)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   27.9 kDa
 
UniProt   P28741 (481-701)
Sequence   FASTA