Limited Proteolysis-Mass Spectrometry Reveals Aging-Associated Changes in Cerebrospinal Fluid Protein Abundances and Structures

Nat Aging. 2022 May;2(5):379-388. doi: 10.1038/s43587-022-00196-x. Epub 2022 Apr 11.

Abstract

Cerebrospinal fluid (CSF) proteins and their structures have been implicated repeatedly in aging and neurodegenerative diseases. Limited proteolysis-mass spectrometry (LiP-MS) is a method that enables proteome-wide screening for changes in both protein abundance and structure. To screen for novel aging-associated changes in the CSF proteome, we performed LiP-MS on CSF from young and old mice with a modified analysis pipeline. We found 38 protein groups change in abundance with aging, most dominantly immunoglobulins of the IgM subclass. We discovered six high-confidence candidates that appeared to change in structure with aging, of which Kng1, Itih2, Lp-PLA2, and 14-3-3 proteins have binding partners or proteoforms known previously to change in the brain with Alzheimer's disease. Intriguingly, using orthogonal validation by Western blot we found the LiP-MS hit Cd5l forms a covalent complex with IgM in mouse and human CSF whose abundance increases with aging. SOMAmer probe signals for all six LiP-MS hits in human CSF, especially 14-3-3 proteins, significantly associate with several clinical features relevant to cognitive function and neurodegeneration. Together, our findings show that LiP-MS can uncover age-related structural changes in CSF with relevance to neurodegeneration.

Publication types

  • Research Support, N.I.H., Extramural
  • Letter
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 14-3-3 Proteins / metabolism
  • Aging
  • Animals
  • Biomarkers / cerebrospinal fluid
  • Cerebrospinal Fluid Proteins* / analysis
  • Humans
  • Immunoglobulin M / metabolism
  • Mice
  • Proteolysis
  • Proteome / analysis
  • Tandem Mass Spectrometry* / methods

Substances

  • Cerebrospinal Fluid Proteins
  • Proteome
  • Biomarkers
  • 14-3-3 Proteins
  • Immunoglobulin M