Pure Appl. Chem., 2012, Vol. 84, No. 9, pp. 1907-1918
Published online 2012-07-30
Balancing life with glycoconjugates: Monitoring unfolded protein response-mediated anti-angiogenic action of tunicamycin by Raman spectroscopy
Asparagine-linked protein glycosylation is a hallmark for glycoprotein structure and function. Its impairment by tunicamycin [a competitive inhibitor
-acetylglucos-aminyl 1-phosphate transferase (GPT)] has been known to inhibit neo-vascularization (i.e., angiogenesis) in humanized breast tumor due to an induction of endoplasmic reticulum (ER) stress-mediated unfolded protein response (UPR). The studies presented here demonstrate that (i) tunicamycin inhibits capillary endothelial cell proliferation in a dose
-dependent manner; (ii) treated cells are incapable of forming colonies upon its withdrawal; and (iii) tunicamycin treatment causes nuclear fragmentation
. Tunicamycin-induced ER stress-mediated UPR event in these cells was studied with the aid of Raman spectroscopy
, in particular, the interpretation of bands at 1672, 1684, and 1694 cm–1
, which are characteristics of proteins
and originate from C=O stretching vibrations of mono-substituted amides
. In tunicamycin-treated cells, these bands decreased in area as follows: at 1672 cm–1
by 41.85 % at 3 h and 55.39 % at 12 h; at 1684 cm–1
by 20.63 % at 3 h and 40.08 % at 12 h; and also at 1994 cm–1
by 33.33 % at 3 h and 32.92 % at 12 h, respectively. Thus, in the presence of tunicamycin, newly synthesized protein chains fail to arrange properly into their final secondary and/or tertiary structures, and the random coils they form had undergone further degradation.
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