5C46

Crystal structure of an engineered construct of phosphatidylinositol 4 kinase III beta in complex with GTP gamma S loaded Rab11

Summary for 5C46

• Entry DOI: 10.2210/pdb5c46/pdb
• Descriptor: Phosphatidylinositol 4-kinase beta, Ras-related protein Rab-11A, SULFATE ION, ... (6 entities in total)
• Functional Keywords: protein-protein complex, lipid kinase, gtpase complex, transferase-signaling protein complex, transferase/signaling protein
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 2
• Total formula weight: 86131.33

Authors

Burke, J.E.,Fowler, M.L. (deposition date: 2015-06-17, release date: 2016-01-20, Last modification date: 2023-09-27)

Primary citation

Fowler, M.L.,McPhail, J.A.,Jenkins, M.L.,Masson, G.R.,Rutaganira, F.U.,Shokat, K.M.,Williams, R.L.,Burke, J.E.
Using hydrogen deuterium exchange mass spectrometry to engineer optimized constructs for crystallization of protein complexes: Case study of PI4KIII beta with Rab11.
Protein Sci., 25:826-839, 2016

PubMed Abstract: The ability of proteins to bind and interact with protein partners plays fundamental roles in many cellular contexts. X-ray crystallography has been a powerful approach to understand protein-protein interactions; however, a challenge in the crystallization of proteins and their complexes is the presence of intrinsically disordered regions. In this article, we describe an application of hydrogen deuterium exchange mass spectrometry (HDX-MS) to identify dynamic regions within type III phosphatidylinositol 4 kinase beta (PI4KIIIβ) in complex with the GTPase Rab11. This information was then used to design deletions that allowed for the production of diffraction quality crystals. Importantly, we also used HDX-MS to verify that the new construct was properly folded, consistent with it being catalytically and functionally active. Structures of PI4KIIIβ in an Apo state and bound to the potent inhibitor BQR695 in complex with both GTPγS and GDP loaded Rab11 were determined. This hybrid HDX-MS/crystallographic strategy revealed novel aspects of the PI4KIIIβ-Rab11 complex, as well as the molecular mechanism of potency of a PI4K specific inhibitor (BQR695). This approach is widely applicable to protein-protein complexes, and is an excellent strategy to optimize constructs for high-resolution structural approaches.

PubMed: 26756197
DOI: 10.1002/pro.2879

Experimental method

X-RAY DIFFRACTION (2.65 Å)