To correlate TIP150 binding to the plus-end tracking of MCAK, we assessed the distribution profiles of MCAK and TIP150 in HeLa cells expressing Flag TIP150 and GFP MCAK deletion mutants.
To date, there have been many crystal structures of the DC-SIGN CRD binding to both oligomannose and LewisX ligands, as well as some showing binding to the CRD plus stalk fragments, that are oligomeric.
Decoration of the abnormal cortical structures by EB1 can be explained by recent studies that show EB1 binding to the lattice as well as the plus ends of growing and shrinking microtubules in the cell periphery (Currie et al., 2011); we also detect EB1 lattice binding to cortical microtubules in wild-type oocytes, making them appear as faint wisplike filaments.
SPR analysis yielded equilibrium dissociation constants for TnC (plus Ca2+) binding to the C-terminal TnI regulatory peptides TnI96 131 and TnI96 139 of 89 nM and 58 nM, respectively.
This assumption can be made due to the positive charge on the amine groups of the polymer backbone, binding to the -COOHs thus rendering them neutral, plus and unbound amine groups which will still hold their positive charge.
As a positive control, we measured the association of phosphorylated ER-α, whose binding to the BRCA1 oligonucleotide was reduced upon cotreatment with estrogen plus PD98059.
α-SNAP showed strong binding to the CAD domain of Stim1 and faint binding to the CAD plus CC1 domain.
Radioactivity (i.e. GTPγS binding to the brain extracts) was measured using MultiLex melt on scintillator sheets (Perkin Elmer) in a liquid scintillation counter (1450 Microbeta Plus; Wallac).
In budding yeast, dynein follows the plus end of the growing microtubule in a Bik1/CLIP (cytoplasmic linker protein -170- and protein -170-endent mander, as a result of transport by the kinesin Kip2 along the microtubule in the direction of the plus end, or as a consequence of direct binding fromannercytoplasm to the plus end [ 33, 34].
This suggests that wild-type dynein can resist its own transport to the plus end through its microtubule-binding domain.
