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Crystallization and preliminary X‐ray crystallographic analysis of the C‐terminal domain of guanylate kinase‐associated protein from Rattus norvegicus
Author(s) -
Tong Junsen,
Yang Huiseon,
Im Young Jun
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x1401187x
Subject(s) - crystallization , crystallography , terminal (telecommunication) , domain (mathematical analysis) , guanylate kinase , materials science , chemistry , biochemistry , computer science , mathematics , membrane protein , organic chemistry , membrane , mathematical analysis , telecommunications
Guanylate kinase‐associated protein (GKAP) is a scaffolding protein that plays a role in protein–protein interactions at the synaptic junction such as linking the NMDA receptor–PSD‐95 complex to the Shank–Homer complex. In this study, the C‐terminal helical domain of GKAP from Rattus norvegicus was purified and crystallized by the vapour‐diffusion method. To improve the diffraction quality of the GKAP crystals, a flexible loop in GKAP was truncated and an MBP (maltose‐binding protein)‐GKAP fusion was constructed in which the last C‐terminal helix of MBP is fused to the N‐terminus of the GKAP domain. The MBP‐GKAP crystals diffracted to 2.0 Å resolution using synchrotron radiation. The crystal was orthorhombic, belonging to space group P 2 1 2 1 2, with unit‐cell parameters a = 99.1, b = 158.7, c = 65.5 Å. The Matthews coefficient was determined to be 2.44 Å 3 Da −1 (solvent content 49.5%) with two molecules in the asymmetric unit. Initial attempts to solve the structure by molecular replacement using the MBP structure were successful.