Advanced search
Start date
Betweenand
(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

X-ray crystallography and NMR studies of domain-swapped canecystatin-1

Full text
Author(s):
Valadares, Napoleao F. [1] ; de Oliveira-Silva, Rodrigo [1] ; Cavini, Italo A. [1] ; Marques, Ivo de Almeida [2] ; Pereira, Humberto D'Muniz [1] ; Soares-Costa, Andrea [3] ; Henrique-Silva, Flavio [3] ; Kalbitzer, Hans R. [4] ; Munte, Claudia E. [1] ; Garratt, Richard C. [1]
Total Authors: 10
Affiliation:
[1] Univ Sao Paulo, Phys Inst Sao Carlos, Ctr Struct Mol Biotechnol, Dept Phys & Informat, BR-13566570 Sao Carlos, SP - Brazil
[2] Univ Fed Goias, Inst Phys, Goiania, Go - Brazil
[3] Univ Fed Sao Carlos, Dept Genet & Evolut, Mol Biol Lab, BR-13560 Sao Carlos, SP - Brazil
[4] Univ Regensburg, Inst Biophys & Phys Biochem, D-93053 Regensburg - Germany
Total Affiliations: 4
Document type: Journal article
Source: FEBS Journal; v. 280, n. 4, p. 1028-1038, FEB 2013.
Web of Science Citations: 9
Abstract

The three-dimensional structure of canecystatin-1, a potent inhibitor of cysteine proteases from sugarcane (Saccharumofficinarum), has been solved in two different crystal forms. In both cases, it is seen to exist as a domain-swapped dimer, the first such observation for a cystatin of plant origin. Size exclusion chromatography and multidimensional NMR spectroscopy show the dimer to be the dominant species in solution, despite the presence of a measurable quantity of monomer undergoing slow exchange. The latter is believed to be the active species, whereas the domain-swapped dimer is presumably inactive, as its first inhibitory loop has been extended to form part of a long -strand that forms a double-helical coiled coil with its partner from the other monomer. A similar structure is observed in human cystatinC, but the spatial disposition of the two lobes of the dimer is rather different. Dimerization is presumably a mechanism by which canecystatin-1 can be kept inactive within the plant, avoiding the inhibition of endogenous proteases. The structure described here provides a platform for the rational design of specific cysteine protease inhibitors for biotechnological applications. Database The coordinates and structure factors have been deposited in the Protein Data Bank under the accession codes 3UL5 and 3UL6. Structured digital abstract Canecystatin-1andCanecystatin-1bindbymolecular sieving(View Interaction:1,2) Canecystatin-1andCanecystatin-1bindbynuclear magnetic resonance(View interaction) Canecystatin-1andCanecystatin-1bindbydynamic light scattering(View interaction) Canecystatin-1andCanecystatin-1bindbyx-ray crystallography(View interaction) (AU)

FAPESP's process: 10/09100-0 - Structural Changes in Hybrid Cystatins Analyzed by High-Resolution Nuclear Magnetic Resonance Spectroscopy
Grantee:Italo Augusto Cavini
Support type: Scholarships in Brazil - Scientific Initiation