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Identification of an Intrinsic Determinant Critical for Maspin Subcellular Localization and Function
Maspin, a multifaceted tumor suppressor, belongs to the serine protease inhibitor superfamily, but only inhibits serine protease-like enzymes such as histone deacetylase 1 (HDAC1). Maspin is specifically expressed in epithelial cells and it is differentially regulated during tumor progression. A new...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3837015/ https://www.ncbi.nlm.nih.gov/pubmed/24278104 http://dx.doi.org/10.1371/journal.pone.0074502 |
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author | Dzinic, Sijana H. Kaplun, Alexander Li, Xiaohua Bernardo, Margarida Meng, Yonghong Dean, Ivory Krass, David Stemmer, Paul Shin, Namhee Lonardo, Fulvio Sheng, Shijie |
author_facet | Dzinic, Sijana H. Kaplun, Alexander Li, Xiaohua Bernardo, Margarida Meng, Yonghong Dean, Ivory Krass, David Stemmer, Paul Shin, Namhee Lonardo, Fulvio Sheng, Shijie |
author_sort | Dzinic, Sijana H. |
collection | PubMed |
description | Maspin, a multifaceted tumor suppressor, belongs to the serine protease inhibitor superfamily, but only inhibits serine protease-like enzymes such as histone deacetylase 1 (HDAC1). Maspin is specifically expressed in epithelial cells and it is differentially regulated during tumor progression. A new emerging consensus suggests that a shift in maspin subcellular localization from the nucleus to the cytoplasm stratifies with poor cancer prognosis. In the current study, we employed a rational mutagenesis approach and showed that maspin reactive center loop (RCL) and its neighboring sequence are critical for maspin stability. Further, when expressed in multiple tumor cell lines, single point mutation of Aspartate(346) (D(346)) to Glutamate (E(346)), maspin(D346E), was predominantly nuclear, whereas wild type maspin (maspin(WT)) was both cytoplasmic and nuclear. Evidence from cellular fractionation followed by immunological and proteomic protein identification, combined with the evidence from fluorescent imaging of endogenous proteins, fluorescent protein fusion constructs, as well as bimolecular fluorescence complementation (BiFC) showed that the increased nuclear enrichment of maspin(D346E) was, at least in part, due to its increased affinity to HDAC1. Maspin(D346E) was also more potent than maspin(WT) as an HDAC inhibitor. Taken together, our evidence demonstrates that D(346) is a critical cis-element in maspin sequence that determines the molecular context and subcellular localization of maspin. A mechanistic model derived from our evidence suggests a new window of opportunity for the development of maspin-based biologically competent HDAC inhibitors for cancer treatment. |
format | Online Article Text |
id | pubmed-3837015 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-38370152013-11-25 Identification of an Intrinsic Determinant Critical for Maspin Subcellular Localization and Function Dzinic, Sijana H. Kaplun, Alexander Li, Xiaohua Bernardo, Margarida Meng, Yonghong Dean, Ivory Krass, David Stemmer, Paul Shin, Namhee Lonardo, Fulvio Sheng, Shijie PLoS One Research Article Maspin, a multifaceted tumor suppressor, belongs to the serine protease inhibitor superfamily, but only inhibits serine protease-like enzymes such as histone deacetylase 1 (HDAC1). Maspin is specifically expressed in epithelial cells and it is differentially regulated during tumor progression. A new emerging consensus suggests that a shift in maspin subcellular localization from the nucleus to the cytoplasm stratifies with poor cancer prognosis. In the current study, we employed a rational mutagenesis approach and showed that maspin reactive center loop (RCL) and its neighboring sequence are critical for maspin stability. Further, when expressed in multiple tumor cell lines, single point mutation of Aspartate(346) (D(346)) to Glutamate (E(346)), maspin(D346E), was predominantly nuclear, whereas wild type maspin (maspin(WT)) was both cytoplasmic and nuclear. Evidence from cellular fractionation followed by immunological and proteomic protein identification, combined with the evidence from fluorescent imaging of endogenous proteins, fluorescent protein fusion constructs, as well as bimolecular fluorescence complementation (BiFC) showed that the increased nuclear enrichment of maspin(D346E) was, at least in part, due to its increased affinity to HDAC1. Maspin(D346E) was also more potent than maspin(WT) as an HDAC inhibitor. Taken together, our evidence demonstrates that D(346) is a critical cis-element in maspin sequence that determines the molecular context and subcellular localization of maspin. A mechanistic model derived from our evidence suggests a new window of opportunity for the development of maspin-based biologically competent HDAC inhibitors for cancer treatment. Public Library of Science 2013-11-21 /pmc/articles/PMC3837015/ /pubmed/24278104 http://dx.doi.org/10.1371/journal.pone.0074502 Text en © 2013 Dzinic et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Dzinic, Sijana H. Kaplun, Alexander Li, Xiaohua Bernardo, Margarida Meng, Yonghong Dean, Ivory Krass, David Stemmer, Paul Shin, Namhee Lonardo, Fulvio Sheng, Shijie Identification of an Intrinsic Determinant Critical for Maspin Subcellular Localization and Function |
title | Identification of an Intrinsic Determinant Critical for Maspin Subcellular Localization and Function |
title_full | Identification of an Intrinsic Determinant Critical for Maspin Subcellular Localization and Function |
title_fullStr | Identification of an Intrinsic Determinant Critical for Maspin Subcellular Localization and Function |
title_full_unstemmed | Identification of an Intrinsic Determinant Critical for Maspin Subcellular Localization and Function |
title_short | Identification of an Intrinsic Determinant Critical for Maspin Subcellular Localization and Function |
title_sort | identification of an intrinsic determinant critical for maspin subcellular localization and function |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3837015/ https://www.ncbi.nlm.nih.gov/pubmed/24278104 http://dx.doi.org/10.1371/journal.pone.0074502 |
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