Cautionary tail: the presence of an N-terminal tag on dynein light-chain Roadblock/LC7 affects its interaction with a functional partner.
As part of structural investigations of components of the molecular motor, dynein, we prepared the light chain, Robl1_mouse, with and without an N-terminal His-tag. We found that the His-tag introduced a spurious binding site for a second protein, IC74. We propose a molecular mechanism for functional interference by the His-tag.
Song
Jikui
J
Center for Eukaryotic Structural Genomics, Department of Biochemistry, University of Wisconsin-Madison, WI 53706-1544, USA.
Markley
John L
JL
D000818:Animals; D001665:Binding Sites; D004398:Dyneins / Q000737:chemistry* / Q000378:metabolism* / Q000502:physiology; D006639:Histidine / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000378:metabolism* / Q000502:physiology
D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D006639:Histidine; D004398:Dyneins; C520623:roadblock 1 protein, mouse