5LNC

Structure of SPX domain of the yeast inorganic polyphophate polymerase Vtc4 crystallized by carrier-driven crystallization in fusion with the macro domain of human histone macroH2A1.1

Summary for 5LNC

• Entry DOI: 10.2210/pdb5lnc/pdb
• Related: 5IIT
• Descriptor: Vacuolar transporter chaperone 4,Core histone macro-H2A.1 (1 entity in total)
• Functional Keywords: macro domain, spx domain, inositol polyphosphate binding, carrier-driven crystallization, hydrolase
• Biological source: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast); More
• Cellular location: Nucleus : O75367
• Total number of polymer chains: 2
• Total formula weight: 84469.87

Authors

Wild, R.,Hothorn, M. (deposition date: 2016-08-03, release date: 2016-11-09, Last modification date: 2024-01-10)

Primary citation

Wild, R.,Hothorn, M.
The macro domain as fusion tag for carrier-driven crystallization.
Protein Sci., 26:365-374, 2017

PubMed Abstract: Obtaining well-ordered crystals remains a significant challenge in protein X-ray crystallography. Carrier-driven crystallization can facilitate crystal formation and structure solution of difficult target proteins. We obtained crystals of the small and highly flexible SPX domain from the yeast vacuolar transporter chaperone 4 (Vtc4) when fused to a C-terminal, non-cleavable macro tag derived from human histone macroH2A1.1. Initial crystals diffracted to 3.3 Å resolution. Reductive protein methylation of the fusion protein yielded a new crystal form diffracting to 2.1 Å. The structures were solved by molecular replacement, using isolated macro domain structures as search models. Our findings suggest that macro domain tags can be employed in recombinant protein expression in E. coli, and in carrier-driven crystallization.

PubMed: 27774698
DOI: 10.1002/pro.3073

Experimental method

X-RAY DIFFRACTION (3.29 Å)