Effects of mutated pregnancy-associated plasma protein-A on atherosclerotic lesion development in mice

Henning B. Boldt, Laurie K. Bale, Zachary T. Resch, Claus Oxvig, Michael T. Overgaard, Cheryl A Conover

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Pregnancy-associated plasma protein-A (PAPP-A) is a large multidomain metalloprotease involved in cleavage of IGF binding protein (IGFBP)-4 and -5 thereby causing release of bioactive IGF. Individual domains of PAPP-A have been characterized in vitro, including the metzincin proteolytic domain important for IGFBP proteolytic activity, short consensus repeats critical for cell surface association, and Lin-12/Notch repeat module demonstrated to determine IGFBP substrate specificity. To test the hypothesis that specific cleavage of IGFBP-4 by PAPP-A in close proximity to the cell surface is required for development of lesions in a murine model of atherosclerosis, the following PAPP-A transgenic (Tg) mice were generated: TgE483A, which lacks all PAPP-A proteolytic activity; TgD1499A, which selectively lacks proteolytic activity against IGFBP-4; and TgK1296A/K1316A, in which cell surface binding is compromised. Following cross-breeding with apolipoprotein E (ApoE) knockout (KO) mice, ApoE KO/Tg mice were fed a high-fat diet to promote aortic lesion development. Lesion area was increased 2-fold in aortas from ApoE KO/Tg wild-type compared with ApoE KO mice (P < 0.001). However, there was no significant increase in the lesion area in any of the ApoE KO/Tg mutant mice. We conclude that PAPP-A proteolytic activity is required for the lesion-promoting effect of PAPP-A and that its specificity must be directed against IGFBP-4. Furthermore, our data demonstrate that cleavage of IGFBP-4 at a distance from the cell surface, and hence from the IGF receptor, is not effective in promoting the development of the atherosclerotic lesions. Thus, PAPP-A exerts its effect while bound to the cell surface in vivo.

Original languageEnglish (US)
Pages (from-to)246-252
Number of pages7
JournalEndocrinology
Volume154
Issue number1
DOIs
StatePublished - Jan 1 2013

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Pregnancy-Associated Plasma Protein-A
Insulin-Like Growth Factor Binding Protein 4
Apolipoproteins E
Knockout Mice
Transgenic Mice
Insulin-Like Growth Factor Binding Proteins
Insulin-Like Growth Factor Binding Protein 5
High Fat Diet
Metalloproteases
Substrate Specificity
Breeding
Aorta
Atherosclerosis

ASJC Scopus subject areas

  • Endocrinology

Cite this

Effects of mutated pregnancy-associated plasma protein-A on atherosclerotic lesion development in mice. / Boldt, Henning B.; Bale, Laurie K.; Resch, Zachary T.; Oxvig, Claus; Overgaard, Michael T.; Conover, Cheryl A.

In: Endocrinology, Vol. 154, No. 1, 01.01.2013, p. 246-252.

Research output: Contribution to journalArticle

Boldt, Henning B. ; Bale, Laurie K. ; Resch, Zachary T. ; Oxvig, Claus ; Overgaard, Michael T. ; Conover, Cheryl A. / Effects of mutated pregnancy-associated plasma protein-A on atherosclerotic lesion development in mice. In: Endocrinology. 2013 ; Vol. 154, No. 1. pp. 246-252.
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abstract = "Pregnancy-associated plasma protein-A (PAPP-A) is a large multidomain metalloprotease involved in cleavage of IGF binding protein (IGFBP)-4 and -5 thereby causing release of bioactive IGF. Individual domains of PAPP-A have been characterized in vitro, including the metzincin proteolytic domain important for IGFBP proteolytic activity, short consensus repeats critical for cell surface association, and Lin-12/Notch repeat module demonstrated to determine IGFBP substrate specificity. To test the hypothesis that specific cleavage of IGFBP-4 by PAPP-A in close proximity to the cell surface is required for development of lesions in a murine model of atherosclerosis, the following PAPP-A transgenic (Tg) mice were generated: TgE483A, which lacks all PAPP-A proteolytic activity; TgD1499A, which selectively lacks proteolytic activity against IGFBP-4; and TgK1296A/K1316A, in which cell surface binding is compromised. Following cross-breeding with apolipoprotein E (ApoE) knockout (KO) mice, ApoE KO/Tg mice were fed a high-fat diet to promote aortic lesion development. Lesion area was increased 2-fold in aortas from ApoE KO/Tg wild-type compared with ApoE KO mice (P < 0.001). However, there was no significant increase in the lesion area in any of the ApoE KO/Tg mutant mice. We conclude that PAPP-A proteolytic activity is required for the lesion-promoting effect of PAPP-A and that its specificity must be directed against IGFBP-4. Furthermore, our data demonstrate that cleavage of IGFBP-4 at a distance from the cell surface, and hence from the IGF receptor, is not effective in promoting the development of the atherosclerotic lesions. Thus, PAPP-A exerts its effect while bound to the cell surface in vivo.",
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