Molecular basis of ice-binding and cryopreservation activities of type III antifreeze proteins
| Autor: | Jaewang Lee, Hyun Sun Kong, Minjae Kim, Yeo Jin Seo, Joon-Hwa Lee, EonSeon Jin, Jung Ryeol Lee, Seo Ree Choi |
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| Rok vydání: | 2021 |
| Předmět: |
wt
wild-type Gene isoform SP sulfopropyl H-bond hydrogen bond Recrystallization (geology) Mutant Biophysics CPA cryoprotective agent AFP Antifreeze protein DMSO dimethyl sulfoxide IRI ice recrystallization inhibition Biochemistry IBP ice-binding protein Cryopreservation OT ovarian tissue 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Structural Biology Antifreeze protein Genetics DSB double-strand break IBS ice-binding surface TH thermal hysteresis ComputingMethodologies_COMPUTERGRAPHICS 030304 developmental biology nfeAFP notched-fin eelpout AFP 0303 health sciences TUNEL terminal deoxynucleotidyl transferase dUTP nick end labelling Chemistry EG ethylene glycol D-PBS Dulbecco’s phosphate-buffered saline Ice crystallization inhibition NMR Computer Science Applications Isoelectric point Ice binding QAE quaternary-amino-ethyl 030220 oncology & carcinogenesis RT room temperature Thermal hysteresis TP248.13-248.65 DNA Research Article Biotechnology |
| Zdroj: | Computational and Structural Biotechnology Journal, Vol 19, Iss, Pp 897-909 (2021) Computational and Structural Biotechnology Journal |
| ISSN: | 2001-0370 |
| DOI: | 10.1016/j.csbj.2021.01.016 |
| Popis: | Graphical abstract Highlights • The QAE2ACT and SP ACT mutants showed full TH and IRI activities. • Active AFPs effectively preserved intact follicle and prevented DSB damage. • Active AFPs exhibited unique structural feature in the first 310 helix of the IBS. • Unique structure of the IBS determines TH, IRI, and cryopreservation activities. Antifreeze proteins (AFPs) can inhibit the freezing of body fluid at subzero temperatures to promote the survival of various organisms living in polar regions. Type III AFPs are categorized into three subgroups, QAE1, QAE2, and SP isoforms, based on differences in their isoelectric points. We determined the thermal hysteresis (TH), ice recrystallization inhibition (IRI), and cryopreservation activity of three isoforms of the notched-fin eelpout AFP and their mutant constructs and characterized their structural and dynamic features using NMR. The QAE1 isoform is the most active among the three classes of III AFP isoforms, and the mutants of inactive QAE2 and SP isoforms, QAE2ACT and SPACT, displayed the full TH and IRI activities with resepect to QAE1 isoform. Cryopreservation studies using mouse ovarian tissue revealed that the QAE1 isoform and the active mutants, QAE2ACT and SPACT, more effectively preserved intact follicle morphology and prevented DNA double-strand break damage more efficiently than the inactive isoforms. It was also found that all active AFPs, QAE1, QAE2ACT, and SPACT, formed unique H-bonds with the first 310 helix, an interaction that plays an important role in the formation of anchored clathrate water networks for efficient binding to the primary prism and pyramidal planes of ice crystals, which was disrupted in the inactive isoforms. Our studies provide valuable insights into the molecular mechanism of the TH and IRI activity, as well as the cryopreservation efficiency, of type III AFPs. |
| Databáze: | OpenAIRE |
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