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Proteolytic Characteristics of Cathepsin D Related to the Recognition and Cleavage of Its Target Proteins

Cathepsin D (CD) plays an important role in both biological and pathological processes, although the cleavage characteristics and substrate selection of CD have yet to be fully explored. We employed liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify the CD cleavage sites in bovine...

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Autores principales: Sun, Huiying, Lou, Xiaomin, Shan, Qiang, Zhang, Ju, Zhu, Xu, Zhang, Jia, Wang, Yang, Xie, Yingying, Xu, Ningzhi, Liu, Siqi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3688724/
https://www.ncbi.nlm.nih.gov/pubmed/23840360
http://dx.doi.org/10.1371/journal.pone.0065733
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author Sun, Huiying
Lou, Xiaomin
Shan, Qiang
Zhang, Ju
Zhu, Xu
Zhang, Jia
Wang, Yang
Xie, Yingying
Xu, Ningzhi
Liu, Siqi
author_facet Sun, Huiying
Lou, Xiaomin
Shan, Qiang
Zhang, Ju
Zhu, Xu
Zhang, Jia
Wang, Yang
Xie, Yingying
Xu, Ningzhi
Liu, Siqi
author_sort Sun, Huiying
collection PubMed
description Cathepsin D (CD) plays an important role in both biological and pathological processes, although the cleavage characteristics and substrate selection of CD have yet to be fully explored. We employed liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify the CD cleavage sites in bovine serum albumin (BSA). We found that the hydrophobic residues at P1 were not only a preferential factor for CD cleavage but that the hydrophobicity at P1’ also contributed to CD recognition. The concept of hydrophobic scores of neighbors (HSN) was proposed to describe the hydrophobic microenvironment of CD recognition sites. The survey of CD cleavage characteristics in several proteins suggested that the HSN was a sensitive indicator for judging the favorable sites in peptides for CD cleavage, with HSN values of 0.5–1.0 representing a likely threshold. Ovalbumin (OVA), a protein resistant to CD cleavage in its native state, was easily cleaved by CD after denaturation, and the features of the cleaved peptides were quite similar to those found in BSA, where a higher HSN value indicated greater cleavability. We further conducted two-dimensional gel electrophoresis (2DE) to find more proteins that were insensitive to CD cleavage in CD-knockdown cells. Based on an analysis of secondary and three-dimensional structures, we postulated that intact proteins with a structure consisting of all α-helices would be relatively accessible to CD cleavage.
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spelling pubmed-36887242013-07-09 Proteolytic Characteristics of Cathepsin D Related to the Recognition and Cleavage of Its Target Proteins Sun, Huiying Lou, Xiaomin Shan, Qiang Zhang, Ju Zhu, Xu Zhang, Jia Wang, Yang Xie, Yingying Xu, Ningzhi Liu, Siqi PLoS One Research Article Cathepsin D (CD) plays an important role in both biological and pathological processes, although the cleavage characteristics and substrate selection of CD have yet to be fully explored. We employed liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify the CD cleavage sites in bovine serum albumin (BSA). We found that the hydrophobic residues at P1 were not only a preferential factor for CD cleavage but that the hydrophobicity at P1’ also contributed to CD recognition. The concept of hydrophobic scores of neighbors (HSN) was proposed to describe the hydrophobic microenvironment of CD recognition sites. The survey of CD cleavage characteristics in several proteins suggested that the HSN was a sensitive indicator for judging the favorable sites in peptides for CD cleavage, with HSN values of 0.5–1.0 representing a likely threshold. Ovalbumin (OVA), a protein resistant to CD cleavage in its native state, was easily cleaved by CD after denaturation, and the features of the cleaved peptides were quite similar to those found in BSA, where a higher HSN value indicated greater cleavability. We further conducted two-dimensional gel electrophoresis (2DE) to find more proteins that were insensitive to CD cleavage in CD-knockdown cells. Based on an analysis of secondary and three-dimensional structures, we postulated that intact proteins with a structure consisting of all α-helices would be relatively accessible to CD cleavage. Public Library of Science 2013-06-20 /pmc/articles/PMC3688724/ /pubmed/23840360 http://dx.doi.org/10.1371/journal.pone.0065733 Text en © 2013 Sun 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
Sun, Huiying
Lou, Xiaomin
Shan, Qiang
Zhang, Ju
Zhu, Xu
Zhang, Jia
Wang, Yang
Xie, Yingying
Xu, Ningzhi
Liu, Siqi
Proteolytic Characteristics of Cathepsin D Related to the Recognition and Cleavage of Its Target Proteins
title Proteolytic Characteristics of Cathepsin D Related to the Recognition and Cleavage of Its Target Proteins
title_full Proteolytic Characteristics of Cathepsin D Related to the Recognition and Cleavage of Its Target Proteins
title_fullStr Proteolytic Characteristics of Cathepsin D Related to the Recognition and Cleavage of Its Target Proteins
title_full_unstemmed Proteolytic Characteristics of Cathepsin D Related to the Recognition and Cleavage of Its Target Proteins
title_short Proteolytic Characteristics of Cathepsin D Related to the Recognition and Cleavage of Its Target Proteins
title_sort proteolytic characteristics of cathepsin d related to the recognition and cleavage of its target proteins
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3688724/
https://www.ncbi.nlm.nih.gov/pubmed/23840360
http://dx.doi.org/10.1371/journal.pone.0065733
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