Proteomic identification of caldesmon as a physiological substrate of proprotein convertase 6 in human uterine decidual cells essential for pregnancy establishment

Lynette M. Kilpatrick, Andrew N. Stephens, Belinda M. Hardman, Lois A. Salamonsen, Ying Li, Peter G. Stanton, Guiying Nie

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Proprotein convertase 5/6 (PC6), a member of the proprotein convertase (PC) family, is a critical regulator in the uterus for embryo implantation. In particular, PC6 is essential for the differentiation of uterine stromal fibroblasts into decidual cells (decidualization). Knockdown of PC6 in the mouse uterus leads to complete failure of decidualization and implantation. It has been envisaged that PC6 functions by proteolytically activating a number of important growth factors and other precursor proteins. However, to date, the precise mechanism of PC6 action in the uterus, particularly during decidualization, is unknown. In this study, we aimed to investigate the mechanisms of PC6 action in decidualization by identifying its physiological substrates using a proteomic approach. Primary human endometrial stromal cells were decidualized and treated with or without recombinant human PC6 (rhPC6). The proteins cleaved by rhPC6 were identified by 2-dimensional fluorescent differential gel electrophoresis. The candidate proteins were validated as PC6 substrates by a number of approaches, including determining their coexpression with PC6 in vivo in decidual cells in the human uterus. A total of 18 protein spots were significantly altered by rhPC6 treatment, 8 of which were assigned clear identities by mass spectrometry. One of these was confirmed to be caldesmon, a key protein involved in organizing the actin microfilaments and regulating cytoskeletal structure. This study demonstrates a novel approach to identify PC-regulated proteins of physiological relevance, and provides important insight into the mechanism by which PC6 regulates decidualization.

Original languageEnglish
Pages (from-to)4983-4992
Number of pages10
JournalJournal of Proteome Research
Volume8
Issue number11
DOIs
Publication statusPublished - 6 Nov 2009
Externally publishedYes

Keywords

  • Decidualization
  • PC5/6
  • Pregnancy 2-dimensional gel electrophoresis
  • Proprotein convertase

Cite this

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title = "Proteomic identification of caldesmon as a physiological substrate of proprotein convertase 6 in human uterine decidual cells essential for pregnancy establishment",
abstract = "Proprotein convertase 5/6 (PC6), a member of the proprotein convertase (PC) family, is a critical regulator in the uterus for embryo implantation. In particular, PC6 is essential for the differentiation of uterine stromal fibroblasts into decidual cells (decidualization). Knockdown of PC6 in the mouse uterus leads to complete failure of decidualization and implantation. It has been envisaged that PC6 functions by proteolytically activating a number of important growth factors and other precursor proteins. However, to date, the precise mechanism of PC6 action in the uterus, particularly during decidualization, is unknown. In this study, we aimed to investigate the mechanisms of PC6 action in decidualization by identifying its physiological substrates using a proteomic approach. Primary human endometrial stromal cells were decidualized and treated with or without recombinant human PC6 (rhPC6). The proteins cleaved by rhPC6 were identified by 2-dimensional fluorescent differential gel electrophoresis. The candidate proteins were validated as PC6 substrates by a number of approaches, including determining their coexpression with PC6 in vivo in decidual cells in the human uterus. A total of 18 protein spots were significantly altered by rhPC6 treatment, 8 of which were assigned clear identities by mass spectrometry. One of these was confirmed to be caldesmon, a key protein involved in organizing the actin microfilaments and regulating cytoskeletal structure. This study demonstrates a novel approach to identify PC-regulated proteins of physiological relevance, and provides important insight into the mechanism by which PC6 regulates decidualization.",
keywords = "Decidualization, PC5/6, Pregnancy 2-dimensional gel electrophoresis, Proprotein convertase",
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Proteomic identification of caldesmon as a physiological substrate of proprotein convertase 6 in human uterine decidual cells essential for pregnancy establishment. / Kilpatrick, Lynette M.; Stephens, Andrew N.; Hardman, Belinda M.; Salamonsen, Lois A.; Li, Ying; Stanton, Peter G.; Nie, Guiying.

In: Journal of Proteome Research, Vol. 8, No. 11, 06.11.2009, p. 4983-4992.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Proteomic identification of caldesmon as a physiological substrate of proprotein convertase 6 in human uterine decidual cells essential for pregnancy establishment

AU - Kilpatrick, Lynette M.

AU - Stephens, Andrew N.

AU - Hardman, Belinda M.

AU - Salamonsen, Lois A.

AU - Li, Ying

AU - Stanton, Peter G.

AU - Nie, Guiying

PY - 2009/11/6

Y1 - 2009/11/6

N2 - Proprotein convertase 5/6 (PC6), a member of the proprotein convertase (PC) family, is a critical regulator in the uterus for embryo implantation. In particular, PC6 is essential for the differentiation of uterine stromal fibroblasts into decidual cells (decidualization). Knockdown of PC6 in the mouse uterus leads to complete failure of decidualization and implantation. It has been envisaged that PC6 functions by proteolytically activating a number of important growth factors and other precursor proteins. However, to date, the precise mechanism of PC6 action in the uterus, particularly during decidualization, is unknown. In this study, we aimed to investigate the mechanisms of PC6 action in decidualization by identifying its physiological substrates using a proteomic approach. Primary human endometrial stromal cells were decidualized and treated with or without recombinant human PC6 (rhPC6). The proteins cleaved by rhPC6 were identified by 2-dimensional fluorescent differential gel electrophoresis. The candidate proteins were validated as PC6 substrates by a number of approaches, including determining their coexpression with PC6 in vivo in decidual cells in the human uterus. A total of 18 protein spots were significantly altered by rhPC6 treatment, 8 of which were assigned clear identities by mass spectrometry. One of these was confirmed to be caldesmon, a key protein involved in organizing the actin microfilaments and regulating cytoskeletal structure. This study demonstrates a novel approach to identify PC-regulated proteins of physiological relevance, and provides important insight into the mechanism by which PC6 regulates decidualization.

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KW - Decidualization

KW - PC5/6

KW - Pregnancy 2-dimensional gel electrophoresis

KW - Proprotein convertase

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U2 - 10.1021/pr900381a

DO - 10.1021/pr900381a

M3 - Article

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SP - 4983

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JO - Journal of Proteome Research

JF - Journal of Proteome Research

SN - 1535-3893

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