5CBP

Crystal Structure of Conjoint Pyrococcus furiosus L-asparaginase at 37 degree C

Summary for 5CBP

• Entry DOI: 10.2210/pdb5cbp/pdb
• Descriptor: L-asparaginase, CITRATE ANION, GLYCEROL, ... (6 entities in total)
• Functional Keywords: hydrolase
• Biological source: Pyrococcus furiosus DSM 3638; More
• Total number of polymer chains: 2
• Total formula weight: 34718.13

Authors

Sharma, P.,Yadav, S.P.,Tomar, R.,Kundu, B.,Ashish, F. (deposition date: 2015-07-01, release date: 2016-07-06, Last modification date: 2024-10-23)

Primary citation

Sharma, P.,Tomar, R.,Yadav, S.S.,Badmalia, M.D.,Nath, S.K.,Kundu, B.
Heat induces end to end repetitive association in P. furiosus L-asparaginase which enables its thermophilic property.
Sci Rep, 10:21702-21702, 2020

PubMed Abstract: It remains undeciphered how thermophilic enzymes display enhanced stability at elevated temperatures. Taking L-asparaginase from P. furiosus (PfA) as an example, we combined scattering shapes deduced from small-angle X-ray scattering (SAXS) data at increased temperatures with symmetry mates from crystallographic structures to find that heating caused end-to-end association. The small contact point of self-binding appeared to be enabled by a terminal short β-strand in N-terminal domain, Leu-Val-Val-Asn (LVVN). Interestingly, deletion of this strand led to a defunct enzyme, whereas suplementation of the peptide LVVN to the defunct enzyme restored structural frameworkwith mesophile-type functionality. Crystal structure of the peptide-bound defunct enzyme showed that one peptide ispresent in the same coordinates as in original enzyme, explaining gain-of lost function. A second peptide was seen bound to the protein at a different location suggesting its possible role in substrate-free molecular-association. Overall, we show that the heating induced self-assembly of native shapes of PfA led to an apparent super-stable assembly.

PubMed: 33303914
DOI: 10.1038/s41598-020-78877-z

Experimental method

X-RAY DIFFRACTION (2.358 Å)