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The role of protein - mineral interactions for protein cycling in soil and protein extraction efficiency from soils


EMSL Project ID
48026

Abstract

We intend (1) to improve the ability to detect proteins in physical soil fractions, and (2) to discover preferences of exoenzymes for relevant classes of mineral components in soil. The latter will be tested by reacting two model proteins differing in their isoelectric points and in molecular weight (beta-glucosidase and a manganese peroxidase) with a variety of functionally different mineral and organic sorbents (Fe-oxide, montmorillonite, quartz and commercially available tannic acid (Sigma Aldrich) at neutral and acidic pH. These systems will subsequently be extracted using a recently developed protocol for metaproteomics analysis (SDS-TCA; Chourey et al. 2010). Consecutive mass – spectrometric analysis conducted at EMSL will allow us to (i) determine how extraction efficiency depends on mineral surface properties, as well as on other relevant controls such as protein molecular weight and the interplay between protein isoelectric point and pH of the system. We will also determine (ii) how different minerals vary in their ability to induce total or partial fragmentation of the protein. Finally (iii), we will attempt to characterize any structural change intact proteins may exhibit after desorption, the latter assuming that intact proteins are found in the extract. The proposed research will provide insight on protein-matrix interactions and help to constrain enzymatic controls on the production of bioavailable C in terrestrial systems. At the same time, we expect to make a significant contribution to the improvement of protein extraction protocols for metaproteomics

Project Details

Project type
Exploratory Research
Start Date
2013-06-16
End Date
2013-09-30
Status
Closed

Team

Principal Investigator

Markus Kleber
Institution
Oregon State University

Team Members

Stephany Chacon
Institution
Lawrence Berkeley National Laboratory

Related Publications

Chacon S.S., P.N. Reardon, C.J. Burgess, S.O. Purvine, R.K. Chu, T.R. Clauss, and E.D. Walter, et al. 2019. "Mineral Surfaces as Agents of Environmental Proteolysis: Mechanisms and Controls." Journal of Environmental Science and Technology 53, no. 6:3018–3026. PNNL-SA-141783. doi:10.1021/acs.est.8b05583