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Isolation of Intermediate Filament Proteins from Multiple Mouse Tissues to Study Aging-associated Post-translational Modifications
Published on: May 18, 2017
Alterations in molecular status of plasma fibronectin associated with aging of normal human individuals
Anna Lemańska-Perek1, Małgorzata Pupek, Bożena Polańska
1Department of Chemistry and Immunochemistry, Wrocław University of Medicine, Bujwida 44a, 50-345 Wrocław, Poland. anna.lemanska-perek@am.wroc.pl
Insights
Plasma fibronectin (FN) molecular status changes with age, showing domain alterations and altered glycosylation in older adults. These changes may link to vascular remodeling during aging.
Area of Science:
- Biochemistry
- Gerontology
- Molecular Biology
Background:
- Senescence involves bodily function deterioration, potentially linked to age-related changes in plasma fibronectin (FN) molecular status.
- Understanding these alterations is crucial for comprehending aging processes.
Purpose of the Study:
- To investigate age-dependent changes in plasma fibronectin (FN) molecular status, including domain expression, glycotope modifications, and molecular forms.
- To correlate these FN alterations with the aging process and potential vascular remodeling.
Main Methods:
- Analyzed 127 plasma samples from healthy individuals across various age groups (newborns to 82 years).
- Utilized FN-ELISA, lectin-FN-ELISA, and immunoblotting with domain-specific antibodies and lectins.
Main Results:
- Plasma FN domains (cell-binding, carboxyl-terminal, collagen, heparin, fibrin) showed significant increases with age.
- Glycosylation patterns, indicated by lectin reactivity (Maackia amurensis, Sambucus nigra), changed significantly with age.
- Novel FN bands (280-kDa, 320-kDa) appeared in older adult groups (41-82 years).
Conclusions:
- Age-related alterations in FN molecular status, including domain changes and glycosylation, are evident throughout the lifespan.
- These modifications may stem from altered FN production/degradation and conformational changes, potentially influencing age-related vascular remodeling.
Objectives:
Senescence, progressive deterioration of many bodily functions might be associated with age-dependent alterations of plasma fibronectin (FN) molecular status (i.e., domain, glycotope, and molecular form expressions).
Design And Methods:
FN molecular status was analyzed in 127 plasma samples of healthy individuals in groups of newborns, and subjects aged 3-14, 15-39, 41-59, and 60-82 years by FN-ELISA, lectin-FN-ELISA, and immunoblotting using a set of domain-specific monoclonal antibodies, specific lectins, and monoclonal antibody to FN, respectively.
Results:
During the first four decades of human life the levels of cell-binding-, carboxyl-terminal-, collagen-, heparin-, and fibrin-domains of plasma FN gradually increased. In subjects aged up to 82 years the cell-binding and carboxyl-terminal FN domain concentrations did not change, while the heparin, fibrin, and collagen domains significantly increased. The relative reactivity of plasma FN with Maackia amurensis lectin, specific to α2,3-linked sialic acid, significantly decreased after birth, reaching a stable level in the subsequent life period, whereas with Sambucus nigra lectin, specific to α2,6-linked sialic acid, it significantly decreased in the 60-82 year old group. Moreover, the appearance of 280-kDa and 320-kDa FN bands, absent in young and mature healthy individuals, was found in the groups of 41-59 and 60-82 year olds.
Conclusions:
The alterations of FN molecular status throughout growth, maturation and senescence might be associated not only with disturbances in the balance of FN production rate and degradation, but concomitantly with conformational rearrangements of FN and its engagement in age-related vascular remodeling processes.
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