title: Crystal structures of the tetratricopeptide repeat domains of kinesin light chains: insight into cargo recognition mechanisms.
abstract: Kinesin-1 transports various cargos along the axon by interacting with the cargos through its light chain subunit. Kinesin light chains (KLC) utilize its tetratricopeptide repeat (TPR) domain to interact with over 10 different cargos. Despite a high sequence identity between their TPR domains (87%), KLC1 and KLC2 isoforms exhibit differential binding properties towards some cargos. We determined the structures of human KLC1 and KLC2 tetratricopeptide repeat (TPR) domains using X-ray crystallography and investigated the different mechanisms by which KLCs interact with their cargos. Using isothermal titration calorimetry, we attributed the specific interaction between KLC1 and JNK-interacting protein 1 (JIP1) cargo to residue N343 in the fourth TRP repeat. Structurally, the N343 residue is adjacent to other asparagines and lysines, creating a positively charged polar patch within the groove of the TPR domain. Whereas, KLC2 with the corresponding residue S328 did not interact with JIP1. Based on these finding, we propose that N343 of KLC1 can form "a carboxylate clamp" with its neighboring asparagine to interact with JIP1, similar to that of HSP70/HSP90 organizing protein-1's (HOP1) interaction with heat shock proteins. For the binding of cargos shared by KLC1 and KLC2, we propose a different site located within the groove but not involving N343. We further propose a third binding site on KLC1 which involves a stretch of polar residues along the inter-TPR loops that may form a network of hydrogen bonds to JIP3 and JIP4. Together, these results provide structural insights into possible mechanisms of interaction between KLC TPR domains and various cargo proteins.
authors.affiliation: Structural Genomics Consortium, Toronto, Ontario, Canada.
authors.forename: Haizhong
authors.initials: H
authors.lastname: Zhu
authors.forename: Han Youl
authors.initials: HY
authors.lastname: Lee
authors.forename: Yufeng
authors.initials: Y
authors.lastname: Tong
authors.forename: Bum-Soo
authors.initials: BS
authors.lastname: Hong
authors.forename: Kyung-Phil
authors.initials: KP
authors.lastname: Kim
authors.forename: Yang
authors.initials: Y
authors.lastname: Shen
authors.forename: Kyung Jik
authors.initials: KJ
authors.lastname: Lim
authors.forename: Farrell
authors.initials: F
authors.lastname: Mackenzie
authors.forename: Wolfram
authors.initials: W
authors.lastname: Tempel
authors.forename: Hee-Won
authors.initials: HW
authors.lastname: Park
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D001665:Binding Sites; D018360:Crystallography, X-Ray; D006801:Humans; D016547:Kinesins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C000715688:KLC1 protein, human; C490666:MAPK8IP1 protein, human; D020033:Protein Isoforms; D011994:Recombinant Proteins; D016547:Kinesins