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Open AccessArticle

A Paradigm in Immunochemistry, Revealed by Monoclonal Antibodies to Spatially Distinct Epitopes on Syntenin-1

1
Mechanisms in Cell Biology and Disease Research Group, School of Pharmacy and Medical Sciences, University of South Australia Cancer Research Institute, University of South Australia, Adelaide, SA 5000, Australia
2
Adelaide Medical School and Freemasons Foundation Centre for Men’s Health, University of Adelaide, Adelaide, SA 5000, Australia
3
South Australian Health and Medical Research Institute, Adelaide, SA 5000, Australia
4
Oxidant and Inflammation Biology Group, Chronic Infectious and Inflammatory Diseases Program, School of Health and Biomedical Sciences, RMIT University, Bundoora, VIC 3803, Australia
5
Department of Pharmacology, Infection and Immunity Program, Biomedicine Discovery Institute, Monash University, Clayton, VIC 3800, Australia
6
Department of Histopathology, School of Medicine Trinity College Dublin 8, Ireland
7
Molecular Pathology Laboratory, The Coombe Women and Infants’ University Hospital, Dublin 8, Ireland
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2019, 20(23), 6035; https://doi.org/10.3390/ijms20236035
Received: 13 November 2019 / Revised: 27 November 2019 / Accepted: 29 November 2019 / Published: 29 November 2019
(This article belongs to the Collection Feature Papers in Molecular Biology)
Syntenin-1 is an essential multi-functional adaptor protein, which has multiple roles in membrane trafficking and exosome biogenesis, as well as scaffolding interactions with either the actin cytoskeleton or focal adhesions. However, how this functional multiplicity relates to syntenin-1 distribution in different endosome compartments or other intracellular locations and its underlying involvement in cancer pathogenesis have yet to be fully defined. To help facilitate the investigation of syntenin-1 biology, we developed two specific monoclonal antibodies (Synt-2C6 and Synt-3A11) to spatially distinct linear sequence epitopes on syntenin-1, which were each designed to be unique at the six-amino acid level. These antibodies produced very different intracellular staining patterns, with Synt-2C6 detecting endosomes and Synt-3A11 producing a fibrillar staining pattern suggesting a cytoskeletal localisation. Treatment of cells with Nocodazole altered the intracellular localisation of Synt-3A11, which was consistent with the syntenin-1 protein interacting with microtubules. In prostate tissue biopsies, Synt-3A11 defined atrophy and early-stage prostate cancer, whereas Synt-2C6 only showed minimal interaction with atrophic tissue. This highlights a critical need for site-specific antibodies and a knowledge of their reactivity to define differential protein distributions, interactions and functions, which may differ between normal and malignant cells.
Keywords: syntenin-1; prostate cancer; microtubules; endosomes syntenin-1; prostate cancer; microtubules; endosomes
MDPI and ACS Style

Johnson, I.R.D.; Sorvina, A.; Logan, J.M.; Moore, C.R.; Heatlie, J.K.; Parkinson-Lawrence, E.J.; Selemidis, S.; O’Leary, J.J.; Butler, L.M.; Brooks, D.A. A Paradigm in Immunochemistry, Revealed by Monoclonal Antibodies to Spatially Distinct Epitopes on Syntenin-1. Int. J. Mol. Sci. 2019, 20, 6035.

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