Effect of Aggregation and Molecular Size on the Ice Nucleation Efficiency of Proteins

Overview

Journal Environ Sci Technol

Publisher American Chemical Society

Specialty Environmental Health

Date 2024 Feb 26

PMID 38408303

Authors

Alyssa N Alsante

Daniel C O Thornton

Sarah D Brooks

Affiliations

Soon will be listed here.

Abstract

Aerosol acts as ice-nucleating particles (INPs) by catalyzing the formation of ice crystals in clouds at temperatures above the homogeneous nucleation threshold (-38 °C). In this study, we show that the immersion mode ice nucleation efficiency of the environmentally relevant protein, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), occurs at temperatures between -6.8 and -31.6 °C. Further, we suggest that this range is controlled by the RuBisCO concentration and protein aggregation. The warmest median nucleation temperature (-7.9 ± 0.8 °C) was associated with the highest concentration of RuBisCO (2 × 10 mg mL) and large aggregates with a hydrodynamic diameter of ∼10 nm. We investigated four additional chemically and structurally diverse proteins, plus the tripeptide glutathione, and found that each of them was a less effective INP than RuBisCO. Ice nucleation efficiency of the proteins was independent of the size (molecular weight) for the five proteins investigated in this study. In contrast to previous work, increasing the concentration and degree of aggregation did not universally increase ice nucleation efficiency. RuBisCO was the exception to this generalization, although the underlying molecular mechanism determining why aggregated RuBisCO is such an effective INP remains elusive.

References

Jassey V, Walcker R, Kardol P, Geisen S, Heger T, Lamentowicz M . Contribution of soil algae to the global carbon cycle. New Phytol. 2022; 234(1):64-76. DOI: 10.1111/nph.17950. View

Yang A, Honig B . On the pH dependence of protein stability. J Mol Biol. 1993; 231(2):459-74. DOI: 10.1006/jmbi.1993.1294. View

Stec B, Hehir M, Brennan C, Nolte M, Kantrowitz E . Kinetic and X-ray structural studies of three mutant E. coli alkaline phosphatases: insights into the catalytic mechanism without the nucleophile Ser102. J Mol Biol. 1998; 277(3):647-62. DOI: 10.1006/jmbi.1998.1635. View

Govindarajan A, Lindow S . Size of bacterial ice-nucleation sites measured in situ by radiation inactivation analysis. Proc Natl Acad Sci U S A. 1988; 85(5):1334-8. PMC: 279765. DOI: 10.1073/pnas.85.5.1334. View

Berman H, Westbrook J, Feng Z, Gilliland G, Bhat T, Weissig H . The Protein Data Bank. Nucleic Acids Res. 1999; 28(1):235-42. PMC: 102472. DOI: 10.1093/nar/28.1.235. View

Zhou X, Entwistle A, Zhang H, Jackson A, Mason T, Shimanovich U . Self-Assembly of Amyloid Fibrils That Display Active Enzymes. ChemCatChem. 2015; 6(7):1961-1968. PMC: 4413355. DOI: 10.1002/cctc.201402125. View

Roeters S, Golbek T, Bregnhoj M, Drace T, Alamdari S, Roseboom W . Ice-nucleating proteins are activated by low temperatures to control the structure of interfacial water. Nat Commun. 2021; 12(1):1183. PMC: 7895962. DOI: 10.1038/s41467-021-21349-3. View

Forman H, Zhang H, Rinna A . Glutathione: overview of its protective roles, measurement, and biosynthesis. Mol Aspects Med. 2008; 30(1-2):1-12. PMC: 2696075. DOI: 10.1016/j.mam.2008.08.006. View

Bar-On Y, Milo R . The global mass and average rate of rubisco. Proc Natl Acad Sci U S A. 2019; 116(10):4738-4743. PMC: 6410859. DOI: 10.1073/pnas.1816654116. View

10.

Metskas L, Ortega D, Oltrogge L, Blikstad C, Lovejoy D, Laughlin T . Rubisco forms a lattice inside alpha-carboxysomes. Nat Commun. 2022; 13(1):4863. PMC: 9388693. DOI: 10.1038/s41467-022-32584-7. View

11.

Qiu Y, Hudait A, Molinero V . How Size and Aggregation of Ice-Binding Proteins Control Their Ice Nucleation Efficiency. J Am Chem Soc. 2019; 141(18):7439-7452. DOI: 10.1021/jacs.9b01854. View

12.

Wang W, Nema S, Teagarden D . Protein aggregation--pathways and influencing factors. Int J Pharm. 2010; 390(2):89-99. DOI: 10.1016/j.ijpharm.2010.02.025. View

13.

MAKI L, Galyan E, Caldwell D . Ice nucleation induced by pseudomonas syringae. Appl Microbiol. 1974; 28(3):456-9. PMC: 186742. DOI: 10.1128/am.28.3.456-459.1974. View

14.

Stetefeld J, McKenna S, Patel T . Dynamic light scattering: a practical guide and applications in biomedical sciences. Biophys Rev. 2017; 8(4):409-427. PMC: 5425802. DOI: 10.1007/s12551-016-0218-6. View

15.

Pandey R, Usui K, Livingstone R, Fischer S, Pfaendtner J, Backus E . Ice-nucleating bacteria control the order and dynamics of interfacial water. Sci Adv. 2016; 2(4):e1501630. PMC: 4846457. DOI: 10.1126/sciadv.1501630. View

16.

Hudait A, Odendahl N, Qiu Y, Paesani F, Molinero V . Ice-Nucleating and Antifreeze Proteins Recognize Ice through a Diversity of Anchored Clathrate and Ice-like Motifs. J Am Chem Soc. 2018; 140(14):4905-4912. DOI: 10.1021/jacs.8b01246. View

17.

Vlasak J, Ionescu R . Fragmentation of monoclonal antibodies. MAbs. 2011; 3(3):253-63. PMC: 3149706. DOI: 10.4161/mabs.3.3.15608. View

18.

Li Y, Lubchenko V, Vekilov P . The use of dynamic light scattering and brownian microscopy to characterize protein aggregation. Rev Sci Instrum. 2011; 82(5):053106. DOI: 10.1063/1.3592581. View

19.

Schuster J, Mahler H, Joerg S, Huwyler J, Mathaes R . Analytical Challenges Assessing Protein Aggregation and Fragmentation Under Physiologic Conditions. J Pharm Sci. 2021; 110(9):3103-3110. DOI: 10.1016/j.xphs.2021.04.014. View

20.

Orellana M, Matrai P, Leck C, Rauschenberg C, Lee A, Coz E . Marine microgels as a source of cloud condensation nuclei in the high Arctic. Proc Natl Acad Sci U S A. 2011; 108(33):13612-7. PMC: 3158224. DOI: 10.1073/pnas.1102457108. View