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Chip-based sensing for release of unprocessed cell surface proteins in vitro and in serum and its (patho)physiological relevance.

Am. J. Physiol. Endocrinol. Metab. 317, E212-E233 (2019)
Verlagsversion Postprint Forschungsdaten DOI
Open Access Green
To study the possibility that certain components of eukaryotic plasma membranes are released under certain (patho)physiological conditions, a chip-based sensor was developed for the detection of cell surface proteins, which are anchored at the outer leaflet of eukaryotic plasma membranes by a covalently attached glycolipid, exclusively, and might be prone to spontaneous or regulated release on the basis of their amphiphilic character. For this, unprocessed, full-length glycosylphosphatidylinositol-anchored proteins (GPI-AP), together with associated phospholipids, were specifically captured and detected by a chip- and microfluidic channel-based sensor, leading to changes in phase and amplitude of surface acoustic waves (SAW) propagating over the chip surface. Unprocessed GPI-AP in complex with lipids were found to be released from rat adipocyte plasma membranes immobilized on the chip, which was dependent on the flow rate and composition of the buffer stream. The complexes were identified in the incubation medium of primary rat adipocytes, in correlation to the cell size, and in rat as well as human serum. With rats, the measured changes in SAW phase shift, reflecting specific mass/size or amount of the unprocessed GPI-AP in complex with lipids, and SAW amplitude, reflecting their viscoelasticity, enabled the differentiation between the lean and obese (high-fat diet) state, and the normal (Wistar) and hyperinsulinemic (Zucker fatty) as well as hyperinsulinemic hyperglycemic (Zucker diabetic fatty) state. Thus chip-based sensing for complexes of unprocessed GPI-AP and lipids reveals the inherently labile anchorage of GPI-AP at plasma membranes and their susceptibility for release in response to (intrinsic/extrinsic) cues of metabolic relevance and may, therefore, be useful for monitoring of (pre-)diabetic disease states.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Schlagwörter Adipocytes ; Biosensor ; Diabetes ; Glycolipid-anchored Cell Surface Proteins ; Metabolic Stress ; Obesity; Gpi-anchored Proteins; Acoustic-wave; Lipid Rafts; Phospholipase-c; Binding; Model; Acetylcholinesterase; Phosphoinositolglycans; Biomarkers; Complex
ISSN (print) / ISBN 0193-1849
e-ISSN 1522-1555
Quellenangaben Band: 317, Heft: 2, Seiten: E212-E233 Artikelnummer: , Supplement: ,
Verlag American Physiological Society
Verlagsort 9650 Rockville Pike, Bethesda, Md 20814 Usa
Begutachtungsstatus Peer reviewed