3IGD

Crystal structure of Mtu recA intein, splicing domain

Summary for 3IGD

• Entry DOI: 10.2210/pdb3igd/pdb
• Related: 2IN0 3IFJ
• Descriptor: Endonuclease PI-MtuI, ZINC ION (3 entities in total)
• Functional Keywords: mini-mini-intein splicing domain, atp-binding, autocatalytic cleavage, cytoplasm, dna damage, dna recombination, dna repair, dna-binding, endonuclease, hydrolase, intron homing, nuclease, nucleotide-binding, protein splicing, sos response, splicing
• Biological source: Mycobacterium tuberculosis; More
• Total number of polymer chains: 1
• Total formula weight: 15504.97

Authors

Van Roey, P.,Belfort, M. (deposition date: 2009-07-27, release date: 2009-10-06, Last modification date: 2023-11-15)

Primary citation

Hiraga, K.,Soga, I.,Dansereau, J.T.,Pereira, B.,Derbyshire, V.,Du, Z.,Wang, C.,Van Roey, P.,Belfort, G.,Belfort, M.
Selection and structure of hyperactive inteins: peripheral changes relayed to the catalytic center.
J.Mol.Biol., 393:1106-1117, 2009

PubMed Abstract: Inteins are phylogenetically diverse self-splicing proteins that are of great functional, evolutionary, biotechnological, and medical interest. To address the relationship between intein structure and function, particularly with respect to regulating the splicing reaction, and to groom inteins for application, we developed a phage display system to extend current in vivo selection for enhanced intein function to selection in vitro. We thereby isolated inteins that can function under excursions in temperature, pH, and denaturing environment. Remarkably, most mutations mapped to the surface of the intein, remote from the active site. We chose two mutants with enhanced splicing activity for crystallography, one of which was also subjected to NMR analysis. These studies define a "ripple effect", whereby mutations in peripheral non-catalytic residues can cause subtle allosteric changes in the active-site environment in a way that facilitates intein activity. Altered salt-bridge formation and chemical shift changes of the mutant inteins provide a molecular rationale for their phenotypes. These fundamental insights will advance the utility of inteins in chemical biology, biotechnology, and medicine.

PubMed: 19744499
DOI: 10.1016/j.jmb.2009.08.074

Experimental method

X-RAY DIFFRACTION (2.4 Å)