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Deciphering the Intratissue-Specific Collagen PTM Site-Specific Heterogeneity in Human Adrenal Extracellular Matrix.

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This study maps post-translational modifications (PTMs) on collagen in the human adrenal gland. We found site-specific differences in collagen PTMs between adrenal fractions, impacting extracellular matrix structure and function.

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Area of Science:

  • Biochemistry
  • Endocrinology
  • Proteomics

Background:

  • The adrenal gland's extracellular matrix (ECM) composition varies between its outer and inner fractions.
  • Collagen, a major ECM component, undergoes crucial post-translational modifications (PTMs) like hydroxylation and glycosylation.
  • Site-specific characterization of collagen PTMs in the human adrenal gland is currently lacking.

Purpose of the Study:

  • To identify and characterize site-specific collagen PTMs within the human adrenal gland.
  • To compare collagen PTMs between the outer fraction (OF) and inner fraction (IF) of the adrenal gland.
  • To investigate the functional implications of observed PTM heterogeneity.

Main Methods:

  • Utilized an in-house proteomics pipeline for high-throughput identification and quantification of collagen PTMs.
  • Analyzed abundant collagen chains from both OF and IF of the human adrenal gland.
  • Developed a COL1A1 PTM map and quantified site-specific modifications.

Main Results:

  • Identified numerous sites for 4-hydroxyproline (4-HyP), 3-hydroxyproline (3-HyP), hydroxylysine (HyK), galactosyl-hydroxylysine (G-HyK), and glucosyl galactosyl-hydroxylysine (GG-HyK).
  • Observed conserved site-specificity but differing occupancies for collagen PTMs between OF and IF.
  • Revealed microheterogeneity in COL1A1 K862, with higher hydroxylation in OF and higher glycosylation in IF, suggesting altered collagen I cross-linking.
  • Found increased site-specific O-glycosylation, particularly GG-HyK, in collagen-IV within the basement membrane.

Conclusions:

  • This study provides the first comprehensive annotation and quantification of site-specific collagen PTMs in the human adrenal gland.
  • Intratissue heterogeneity in collagen PTMs exists even within a seemingly stable ECM, with distinct patterns in OF and IF.
  • The observed PTM variations, especially in collagen-IV glycosylation, may play a role in adrenal steroid secretion and region-specific functions.