BIOISIS ID :: CAN1AP

apo-state Type III CRISPR ancillary nuclease 1 (can1)

Added: 29 September 2020

Description:

CRISPR ancillary nuclease from T. thermophilus was crystallised in the presence of cyclic tetra-adenylate (cA4). Attempts to crystallise can1 in absence of cA4 failed to produce suitable crystals. The can1 nuclease is upregulated during phage infection and presumably participates in anti-viral defence.

Total datasets deposited with submission: 1

Supporting archive file:

Download: best_apo_models.zip

CNS solve (Brunger) was used to complete the X-ray crystal structure. Subsequent rounds of simulated annealing using distance restraints derived from crystal structure and bounded by P( r )-distribution allowed an exploration of conformational space. Files in archive describe the best fit, contains tbl restraints and model_anneal.inp


Data Set:

SEC SAXS dataset of apo-state CRISPR III can1

Description:

SEC SAXS of can1 was performed using a Shodex KW-402.5 column pre-equilibrated with 3 column volumes of buffer. SEC-SAXS experiments were measured at a flow rate of 0.160 ml per minute with 2 s exposure per frame in a 1.5 mm diameter, 10 μm thick quartz capillary flow cell and a 800 μm squared X-ray beam focused on an Eiger 4 M detector 4 m from the sample. Column performance and instrumentation calibration was checked by measuring a 45 μl sample of BSA injected at 10 mg mL^−1^. In total 45 μl of the apo-state of Can1 was injected (10.4 mg mL^−1^, 140 μM) and frames were collected for a total of 30 min.

Buffer (Matrix):

20 mM Tris pH 7.5 and 150 mM NaCl

Source: synchrotron

Instrument: B21 Diamond Light Source

Deposited files in dataset:

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Real-space Transform: apo_pr.dat dmax :: 95.0

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Download apo_pr.dat
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