Complimentary protein extraction methods increase the identification of the Park Grass Experiment metaproteome

Although the Park Grass Experiment is an important international reference soil for temperate grasslands, it still lacks direct extraction of its metaproteome. The identification these proteins can be crucial to our understanding ecology and major biogeochemical processes. However, protein from a technically fraught process due difficulties with co-extraction humic material lack compatible databases identify proteins. To address issues, we combined two techniques on experiment soil, one based acid removal, namely modified freeze-dry, heat/thaw/phenol/chloroform (HTPC) method another which co-extracts material, established surfactant method. A broad range were identified by matching mass spectra extracted against tailored proteome database. These mainly in categories “protein metabolism”, “membrane transport”, “carbohydrate “respiration” “ribosomal nitrogen cycle” proteins, enabling reconstitution specific processes grassland soil. Protein annotation using NCBI EBI inferred that dominated Proteobacteria, Actinobacteria, Acidobacteria Firmicutes at phylum level Bradyrhizobium, Rhizobium, Acidobacteria, Streptomyces Pseudolabrys genus level. Further functional enrichment analysis enabled us connect identities regulatory signalling networks key cycles, notably cycle. newly metaproteome thus provides baseline future targeted studies their control built.