Quantitative profiling of microbial communities by de novo metaproteomics

preprint · bioRxiv · 2020

preprint · bioRxiv · 2020. Hugo B.C. Kleikamp et al. Metaproteomics has emerged as one of the most promising approaches for determining the composition and…
Date 2020-08-17
Type preprint
Venue bioRxiv
Publisher openRxiv
Contribution downstream-application
DOI 10.1101/2020.08.16.252924
Citations (OpenAlex) 3

Peer-reviewed version: Database-independent de novo metaproteomics of complex microbial communities (2021-05-01, Cell Systems)

Abstract

Metaproteomics has emerged as one of the most promising approaches for determining the composition and metabolic functions of complete microbial communities. Conventional metaproteomics approaches however, rely on the construction of protein sequence databases and efficient peptide-spectrum matching algorithms. Thereby, very large sequence databases impact on computational efforts and sensitivity. More recently, advanced de novo sequencing strategies–which annotate peptide sequences without the requirement for a database–have become (again) increasingly proposed for proteomics applications. Such approaches would vastly expand many metaproteomics applications by enabling rapid community profiling and by capturing unsequenced community members, which otherwise remain inaccessible for further interpretation. Nevertheless, because of the lack of efficient pipelines and validation procedures, those strategies have only rarely been employed for community proteomics. Here we report on a newly established de novo metaproteomics pipeline which was evaluated for its quantitative performance using synthetic and natural communities. Additionally, we introduce a novel validation strategy and investigate the actual content of community members within community proteomics data.

Authors

  1. Hugo B.C. Kleikamp · Delft University of Technology, University of Antwerp
  2. Mario Pronk · Delft University of Technology
  3. Claudia G. Tugui · Delft University of Technology
  4. Leonor Guedes da Silva · Delft University of Technology
  5. Ben Abbas · Delft University of Technology
  6. Yue Mei Lin · Delft University of Technology
  7. Mark C. M. van Loosdrecht · Delft University of Technology
  8. Martin Pabst · Delft University of Technology

Methods and tools

  • NovoBridge: A database-independent metaproteomics pipeline that maps de novo peptide sequences to taxonomy and function, giving quantitative community profiles without a sample-specific database.

Methods it uses

  • PEAKS: Commercial DP-based de novo

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