Identification, biochemical characterization and crystallization of the central region of human ATG16L1

In this study, the authors aimed to structurally characterize the N-terminal half of ATG16L1. ATG16L111 – 307 in complex with ATG5 has been purified and crystallized in two crystal forms. However, both crystal structures revealed degradation of ATG16L1, resulting in crystals comprising only full-length ATG5 and the ATG5BD of ATG16L1. The structures of ATG5 – ATG5BD in two novel crystal forms are presented, further supporting the previously observed dimerization of ATG5 – ATG16L1. The reported degradation points towards a high instability at the linker region between the ATG5BD and the CCD in ATG16L1. Based on this observation and further biochemical analysis of ATG16L1, a stable 236-amino-acid subfragment comprising residues 72 – 307 of the N-terminal half of ATG16L1, covering the residual, so far structurally uncharacterized region of human ATG16L1, was identified. Here, the identification, purification, biochemical characterization and crystallization of the proteolytically stable ATG16L172 – 307 subfragment are reported.

Source: Acta Crystallographica Section F - September 28, 2017 Category: Biochemistry Authors: Archna, A. Scrima, A. Tags: autophagy ATG16L1 ATG5 ATG5BD coiled coil research communications Source Type: research