There is a lack in understanding about the role of characteristic mixtures of osmolytes in normal body function, and in many disease conditions, including diabetes mellitus and cancer. The long-term goal is to understand the normal and abnormal roles of these osmolytes in humans, and to design strategies for the use of osmolytes in the treatment and prevention of disease. This project's objective is to understand how non-specific (preferential) interactions contribute to the apparent necessity of osmolyte mixtures observed in vivo. It is hypothesized that the requirement for complex osmolyte cocktails is intrinsic to aqueous mixtures of proteins, nucleotide ligands, and osmolytes. The rationale is that this research provides the basis for understanding the role that osmolyte mixtures play in so many important normal and abnormal cellular processes. These include normal tissue function in kidney and brain, and the pathological impact of osmolytes in many diseases. The central hypothesis will be tested by pursuing Specific Aims that address the major reasons why complex osmolyte mixtures may be necessary: disparate impact of osmolytes on proteins and nucleotides, which leads to the necessity of the mixtures;and non-additive action of osmolytes, which leads to complex, non-linear behavior. The impact of osmolytes on protein-nucleotide systems will be quantified through high-throughput methods. Thermal melts and measurements of effective concentrations will be performed on microplates, and conventional approaches will be used to verify the results. The initial model proteins are adenylate kinase, dihydrofolate reductase, and carbonic anhydrase, in conjunction with small nucleotide ligands, including ATP, AMP, NADP, and others. Thirteen osmolytes that are common in humans and throughout nature will be used in binary mixtures. This project is innovative in its idea that pathological osmolyte mixtures are a contributing factor to many deadly diseases. We expect that knowledge of the effects of natural osmolyte cocktails on proteins and nucleotides will allow differentiating between deleterious and beneficial mixtures, and elucidate osmolytes'contributing role in many diseases. Such information would be very valuable in identifying targets for the manipulation of the osmolyte content of cells through medication, supplements, or diet. This project is significant, because it overcomes a major block in understanding cellular pathophysiology, namely the question of what new properties emerge when osmolytes are combined in their natural mixtures with proteins and metabolites, thus opening new opportunities of ameliorating disease. Additional benefits of this kind of research include an enhanced understanding of protein folding, of protein crystallization, and of drug formulation.

Public Health Relevance

This project investigates mixtures of osmolytes (small molecules that naturally occur in humans), and how some of these cocktails can lead to beneficial or detrimental effects on biomolecules. It is expected that these results will facilitate the search for efficient chemoprevention agents against various cancers, and other diseases in which osmolytes play a role. Narrative This project investigates mixtures of osmolytes (small molecules that naturally occur in humans), and how some of these cocktails can lead to beneficial or detrimental effects on biomolecules. It is expected that these results will facilitate the search for efficient chemoprevention agents against various cancers, and other diseases in which osmolytes play a role.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01GM049760-22
Application #
8726415
Study Section
Macromolecular Structure and Function B Study Section (MSFB)
Program Officer
Smith, Ward
Project Start
Project End
Budget Start
Budget End
Support Year
22
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Pennsylvania State University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
City
Hershey
State
PA
Country
United States
Zip Code
17033
Rajarathnam, Krishna; Rosgen, Jorg (2014) Isothermal titration calorimetry of membrane proteins - progress and challenges. Biochim Biophys Acta 1838:69-77
Jackson-Atogi, Ruby; Sinha, Prem Kumar; Rosgen, Jorg (2013) Distinctive solvation patterns make renal osmolytes diverse. Biophys J 105:2166-74
Rosgen, Jorg; Jackson-Atogi, Ruby (2012) Volume exclusion and H-bonding dominate the thermodynamics and solvation of trimethylamine-N-oxide in aqueous urea. J Am Chem Soc 134:3590-7
Holthauzen, Luis Marcelo F; Auton, Matthew; Sinev, Mikhail et al. (2011) Protein stability in the presence of cosolutes. Methods Enzymol 492:61-125
Auton, Matthew; Rosgen, Jorg; Sinev, Mikhail et al. (2011) Osmolyte effects on protein stability and solubility: a balancing act between backbone and side-chains. Biophys Chem 159:90-9
Holthauzen, Luis Marcelo F; Rosgen, Jorg; Bolen, D Wayne (2010) Hydrogen bonding progressively strengthens upon transfer of the protein urea-denatured state to water and protecting osmolytes. Biochemistry 49:1310-8
Street, Timothy O; Krukenberg, Kristin A; Rosgen, Jorg et al. (2010) Osmolyte-induced conformational changes in the Hsp90 molecular chaperone. Protein Sci 19:57-65
Hu, Char Y; Kokubo, Hironori; Lynch, Gillian C et al. (2010) Backbone additivity in the transfer model of protein solvation. Protein Sci 19:1011-22
Schrank, Travis P; Bolen, D Wayne; Hilser, Vincent J (2009) Rational modulation of conformational fluctuations in adenylate kinase reveals a local unfolding mechanism for allostery and functional adaptation in proteins. Proc Natl Acad Sci U S A 106:16984-9
Lim, Woon Ki; Rosgen, Jorg; Englander, S Walter (2009) Urea, but not guanidinium, destabilizes proteins by forming hydrogen bonds to the peptide group. Proc Natl Acad Sci U S A 106:2595-600

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