6SWU

Crystal structure of the TPR domain of KLC1 in complex with an engineered high-affinity cargo peptide.

Summary for 6SWU

• Entry DOI: 10.2210/pdb6swu/pdb
• Descriptor: Kinesin light chain 1,Kinesin light chain 1,TPR domain of kinesin light chain 1 in complex with an engineered high-affinity cargo peptide of sequence TVFTTEDIYEWDDSAI, DI(HYDROXYETHYL)ETHER, 1,2-ETHANEDIOL, ... (4 entities in total)
• Functional Keywords: protein complex, protein engineering, motor protein, cargo recognition
• Biological source: Mus musculus (House mouse); More
• Total number of polymer chains: 6
• Total formula weight: 228275.20

Authors

Chegkazi, M.S.,Steiner, R.A. (deposition date: 2019-09-23, release date: 2020-09-30, Last modification date: 2024-01-24)

Primary citation

Cross, J.A.,Chegkazi, M.S.,Steiner, R.A.,Woolfson, D.N.,Dodding, M.P.
Fragment-linking peptide design yields a high-affinity ligand for microtubule-based transport.
Cell Chem Biol, 28:1347-, 2021

PubMed Abstract: Synthetic peptides are attractive candidates to manipulate protein-protein interactions inside the cell as they mimic natural interactions to compete for binding. However, protein-peptide interactions are often dynamic and weak. A challenge is to design peptides that make improved interactions with the target. Here, we devise a fragment-linking strategy-"mash-up" design-to deliver a high-affinity ligand, KinTag, for the kinesin-1 motor. Using structural insights from natural micromolar-affinity cargo-adaptor ligands, we have identified and combined key binding features in a single, high-affinity ligand. An X-ray crystal structure demonstrates interactions as designed and reveals only a modest increase in interface area. Moreover, when genetically encoded, KinTag promotes transport of lysosomes with higher efficiency than natural sequences, revealing a direct link between motor-adaptor binding affinity and organelle transport. Together, these data demonstrate a fragment-linking strategy for peptide design and its application in a synthetic motor ligand to direct cellular cargo transport.

PubMed: 33838110
DOI: 10.1016/j.chembiol.2021.03.010

Experimental method

X-RAY DIFFRACTION (2.849 Å)