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The anti-apoptotic activity of albumin for endothelium is inhibited by advanced glycation end products restricting intramolecular movement

Cellular & Molecular Biology Letters volume 14, pages 575–586 (2009)Cite this article

Abstract

Human serum albumin (HSA) inhibits endothelial apoptosis in a highly specific manner. CNBr fragmentation greatly increases the effectiveness of this activity, suggesting that this type of protection is mediated by a partially cryptic albumin domain which is transiently exposed by intramolecular movement. Advanced glycation end-product (AGE) formation in HSA greatly reduces its intra-molecular movement. This study aimed to determine if this inhibits the anti-apoptotic activity of HSA, and if such inactivation could be reversed by CNBr fragmentation. HSA-AGE was prepared by incubating HSA with glucose, and assessed using the fructosamine assay, mass spectrometry, SDS-PAGE and fluorometry. Low levels of AGE in the HSA had little effect upon its anti-apoptotic activity, but when the levels of AGE were high and the intra-molecular movement was reduced, endothelial cell survival was also found to be reduced to levels equivalent to those in cultures without HSA or serum (p > 0.001). Survival was restored by the inclusion of native HSA, despite the presence of HSA with high levels of AGE. Also, CNBr fragmentation of otherwise inactive HSA-AGE restored the anti-apoptotic activity for endothelium. Apoptosis was confirmed by DNA gel electrophoresis, transmission electron microscopy and fluorescence-activated cell sorting analysis, and there was no evidence for direct toxicity in the HSA-AGE preparations. The results are consistent with the proposed role of intra-molecular movement in exposing the anti-apoptotic domain in HSA for endothelium. The levels of AGE formation required to inhibit the anti-apoptotic activity of HSA exceeded those reported for diabetes. Nonetheless, the data from this study seems to be the first example of reduced protein function due to AGE-restricted intra-molecular movement.

Abbreviations

AGE:

advanced glycation end product

HSA:

human serum albumin

HUVEC:

human umbilical vein endothelial cells

PBS:

phosphate-buffered saline

SCS:

supplemented calf serum

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Authors and Affiliations

  1. Cellular and Molecular Pathology Research Unit, Department of Oral Pathology and Oral Medicine, Westmead Centre for Oral Health, The University of Sydney, Westmead, NSW, 2145, Australia

    Hans Zoellner, Salman Siddiqui & Elizabeth Kelly

  2. Department of Surgery, Westmead Hospital, Westmead, NSW, 2145, Australia

    Heather Medbury

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  1. Hans Zoellner
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  2. Salman Siddiqui
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  3. Elizabeth Kelly
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Correspondence to Hans Zoellner.

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Zoellner, H., Siddiqui, S., Kelly, E. et al. The anti-apoptotic activity of albumin for endothelium is inhibited by advanced glycation end products restricting intramolecular movement. Cell Mol Biol Lett 14, 575–586 (2009). https://doi.org/10.2478/s11658-009-0021-5

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Cellular & Molecular Biology Letters

ISSN: 1689-1392

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