Structural basis of the correct subunit assembly, aggregation, and intracellular degradation of nylon hydrolase
| Autor: | Dai-ichiro Kato, Young-Ho Lee, Yusuke Tanaka, Naoki Shibata, Yuji Goto, Ikki Takehara, Masahiro Takeo, Seiji Negoro, Yoshiki Higuchi, Keisuke Nagai, Ryo Kinugasa |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Protein subunit Mutant lcsh:Medicine Article Protein Structure Secondary 03 medical and health sciences Bacterial Proteins Aminohydrolases Hydrolase lcsh:Science Thermostability chemistry.chemical_classification Multidisciplinary lcsh:R Amino acid Nylons 030104 developmental biology Enzyme chemistry Biophysics lcsh:Q Protein quaternary structure Peptides Dimerization Intracellular |
| Zdroj: | Scientific Reports Scientific Reports, Vol 8, Iss 1, Pp 1-16 (2018) |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/s41598-018-27860-w |
| Popis: | Nylon hydrolase (NylC) is initially expressed as an inactive precursor (36 kDa). The precursor is cleaved autocatalytically at Asn266/Thr267 to generate an active enzyme composed of an α subunit (27 kDa) and a β subunit (9 kDa). Four αβ heterodimers (molecules A-D) form a doughnut-shaped quaternary structure. In this study, the thermostability of the parental NylC was altered by amino acid substitutions located at the A/D interface (D122G/H130Y/D36A/L137A) or the A/B interface (E263Q) and spanned a range of 47 °C. Considering structural, biophysical, and biochemical analyses, we discuss the structural basis of the stability of nylon hydrolase. From the analytical centrifugation data obtained regarding the various mutant enzymes, we conclude that the assembly of the monomeric units is dynamically altered by the mutations. Finally, we propose a model that can predict whether the fate of the nascent polypeptide will be correct subunit assembly, inappropriate protein-protein interactions causing aggregation, or intracellular degradation of the polypeptide. |
| Databáze: | OpenAIRE |
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