KIF22 Motor Domain (5-378) His-Protein: wild-type (Human recombinant)
Measurement of microtubule-activated ATPase assays
Identification/characterization of proteins or small molecules that affect motor ATPase activity.
Identification/characterization of proteins or small molecules that affect motor/microtubule interactions.
The wild-type human motor domain of the kinesin-like KIF22 protein (also known as kinesin-like DNA binding protein, KID) has been produced in a bacterial expression system. The recombinant protein contains a 6x His-tag at the amino terminal-end and amino acids 5-378. The molecular weight of KIF22 is approximately 43 kDa and it is supplied as a white lyophilized powder.
Protein purity is determined by scanning densitometry of Coomassie Blue-stained protein on a 4-20% polyacrylamide gradient gel. Kif22 5-378 protein was determined to be 90% pure. (see Figure 1 for an example).
Figure 1. KIF22 5-378 Protein Purity Determination. A 10 µg sample of recombinant KIF22 protein (molecular weight approx. 43 kDa) was separated by electrophoresis in a 4-20% SDS-PAGE system and stained with Coomassie Blue. Protein quantitation was determined using the Precision Red Protein Assay Reagent (Cat. # ADV02). Seeblue Pre-stained molecular weight markers are from Invitrogen.
Figure 2: KIF22 (0.4 µg) ATPase activity was measured (n=4) at an absorbance of 360 nm on an iD5 multi-mode micro-plate reader (Molecular devices) over 30 minutes at room temper-ature. The ATPase activity was started when 0.6 mM of ATP was added 3 minutes into the kinetic run. Control reactions (n=3) were carried out in the absence of KIF22 motor protein.