In:
IUBMB Life, Wiley, Vol. 71, No. 3 ( 2019-03), p. 364-375
Abstract:
Emerging evidence on efficient tumor growth regulation by endogenous lectins directs interest to determine on a proof‐of‐principle level the range of information on alterations provided by full‐scale analysis using phosphoproteomics. In our pilot study, we tested galectin‐4 (gal‐4) that is a growth inhibitor for colon cancer cells (CRC), here working with the LS 180 line. In order to cover monitoring of short‐ and long‐term effects stable isotope labeling by amino acids in cell culture‐based quantitative phosphoproteomic analyses were conducted on LS 180 cell preparations collected 1 and 72 h after adding gal‐4 to the culture medium. After short‐term treatment, 981 phosphosites, all of them S/T based, were detected by phosphoproteomics. Changes higher than 1.5‐fold were seen for eight sites in seven proteins. Most affected were the BET1 homolog (BET1), whose level of phosphorylation at S50 was about threefold reduced, and centromere protein F (CENPF), extent of phosphorylation at S3119 doubling in gal‐4‐treated cells. Phosphoproteome analysis after 72 h of treatment revealed marked changes at 33 S/T‐based phosphosites from 29 proteins. Prominent increase of phosphorylation was observed for cofilin‐1 at position S3. Extent of phosphorylation of the glutamine transporter SLC1A5 at position S503 was decreased by a factor of 3. Altered phosphorylation of BET1, CENPF, and cofilin‐1 as well as a significant effect of gal‐4 treatment on glutamine uptake by cells were substantiated by independent methods in the Vaco 432, Colo 205, CX 1, and HCT 116 cell lines. With the example of gal‐4 which functions as a tumor suppressor in CRC cells, we were able to prove that cell surface binding of the lectin not only markedly influences the cell proteome, but also has a bearing on malignancy‐associated intracellular protein phosphorylation . These results underscore the potential of this approach to give further work on elucidating the details of signaling underlying galectin‐triggered growth inhibition a clear direction. © 2018 IUBMB Life, 71(3):364–375, 2019
Type of Medium:
Online Resource
ISSN:
1521-6543
,
1521-6551
Language:
English
Publisher:
Wiley
Publication Date:
2019
detail.hit.zdb_id:
2009952-6
detail.hit.zdb_id:
2485214-4
SSG:
12
