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A corin variant identified in hypertensive patients that alters cytoplasmic tail and reduces cell surface expression and activity.

Zhang Y, Li H, Zhou J, Wang A, Yang J, Wang C, Liu M, Zhou T, Zhu L, Zhang Y, Dong N, Wu Q - Sci Rep (2014)

Bottom Line: Corin is a membrane-bound protease that regulates blood pressure by activating the natriuretic peptides.Analysis of two independent cohorts showed that the variant was preferentially present in hypertensive patients (38/795 or 4.78% vs. 4/632 or 0.63% in normal individuals, p = 4.14E-6).In cell-based studies, the corin variant exhibited poor trafficking in the Golgi, reduced cell surface expression and zymogen activation, and low natriuretic peptide processing activity.

View Article: PubMed Central - PubMed

Affiliation: Cyrus Tang Hematology Center, MOE Engineering Center of Hematological Disease, MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital, and Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China.

ABSTRACT
Corin is a membrane-bound protease that regulates blood pressure by activating the natriuretic peptides. CORIN variants have been associated with hypertension and heart disease in African Americans. In this study, we conducted targeted exome sequencing and identified an insertion variant, c.102_103insA, in exon 1 of the CORIN gene. Analysis of two independent cohorts showed that the variant was preferentially present in hypertensive patients (38/795 or 4.78% vs. 4/632 or 0.63% in normal individuals, p = 4.14E-6). The insertion shifted the reading frame, resulting in a corin variant with a truncated cytoplasmic tail. In cell-based studies, the corin variant exhibited poor trafficking in the Golgi, reduced cell surface expression and zymogen activation, and low natriuretic peptide processing activity. Compared with normal individuals with the wild-type allele, individuals with the variant allele had lower levels of plasma corin [0.59 ± 0.07 ng/mL (n = 25) vs. 0.91 ± 0.02 ng/mL (n = 215), p<0.001] and higher levels of plasma N-terminal pro-atrial natriuretic peptide (NT-pro-ANP) [2.39 ± 3.6 nmol/L (n = 21) vs. 0.87 ± 0.6 nmol/L (n = 48), p = 0.005]. These results indicate that the variant altered corin structure and impaired the natriuretic peptide processing activity in vivo. The results highlight corin defects as an important underlying mechanism in hypertension.

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Expression and activity of the insA variant in HEK293 cells.(A) Western analysis of corin WT, the insA variant, and mutants R801A and S985A protein levels in transfected HEK293 cells. (B) Pro-ANP processing activity of the insA variant. WT and inactive mutants R801A and S985A were positive and negative controls, respectively. (C) Quantitative data of pro-ANP processing activity were from densitometric analysis. The cropped blots are used in the main figures and full-length blots are included in the supplementary information (Supplementary Fig. S1). Data are representative of at least four independent experiments.
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f2: Expression and activity of the insA variant in HEK293 cells.(A) Western analysis of corin WT, the insA variant, and mutants R801A and S985A protein levels in transfected HEK293 cells. (B) Pro-ANP processing activity of the insA variant. WT and inactive mutants R801A and S985A were positive and negative controls, respectively. (C) Quantitative data of pro-ANP processing activity were from densitometric analysis. The cropped blots are used in the main figures and full-length blots are included in the supplementary information (Supplementary Fig. S1). Data are representative of at least four independent experiments.

Mentions: The insertion of an adenine at nucleotide position 102 shifts the reading frame (Fig. 1B), which is expected to prevent corin expression. We made the plasmid, pcDNAinsA, containing the adenine insertion and transfected it in HEK293 cells. WT corin (positive) and inactive mutants R801A and S985A (negative) were used as controls (Fig. 1B). Surprisingly, Western analysis detected the insA variant expression in transfected HEK293 cells (Fig. 2A).


A corin variant identified in hypertensive patients that alters cytoplasmic tail and reduces cell surface expression and activity.

Zhang Y, Li H, Zhou J, Wang A, Yang J, Wang C, Liu M, Zhou T, Zhu L, Zhang Y, Dong N, Wu Q - Sci Rep (2014)

Expression and activity of the insA variant in HEK293 cells.(A) Western analysis of corin WT, the insA variant, and mutants R801A and S985A protein levels in transfected HEK293 cells. (B) Pro-ANP processing activity of the insA variant. WT and inactive mutants R801A and S985A were positive and negative controls, respectively. (C) Quantitative data of pro-ANP processing activity were from densitometric analysis. The cropped blots are used in the main figures and full-length blots are included in the supplementary information (Supplementary Fig. S1). Data are representative of at least four independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4260221&req=5

f2: Expression and activity of the insA variant in HEK293 cells.(A) Western analysis of corin WT, the insA variant, and mutants R801A and S985A protein levels in transfected HEK293 cells. (B) Pro-ANP processing activity of the insA variant. WT and inactive mutants R801A and S985A were positive and negative controls, respectively. (C) Quantitative data of pro-ANP processing activity were from densitometric analysis. The cropped blots are used in the main figures and full-length blots are included in the supplementary information (Supplementary Fig. S1). Data are representative of at least four independent experiments.
Mentions: The insertion of an adenine at nucleotide position 102 shifts the reading frame (Fig. 1B), which is expected to prevent corin expression. We made the plasmid, pcDNAinsA, containing the adenine insertion and transfected it in HEK293 cells. WT corin (positive) and inactive mutants R801A and S985A (negative) were used as controls (Fig. 1B). Surprisingly, Western analysis detected the insA variant expression in transfected HEK293 cells (Fig. 2A).

Bottom Line: Corin is a membrane-bound protease that regulates blood pressure by activating the natriuretic peptides.Analysis of two independent cohorts showed that the variant was preferentially present in hypertensive patients (38/795 or 4.78% vs. 4/632 or 0.63% in normal individuals, p = 4.14E-6).In cell-based studies, the corin variant exhibited poor trafficking in the Golgi, reduced cell surface expression and zymogen activation, and low natriuretic peptide processing activity.

View Article: PubMed Central - PubMed

Affiliation: Cyrus Tang Hematology Center, MOE Engineering Center of Hematological Disease, MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital, and Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China.

ABSTRACT
Corin is a membrane-bound protease that regulates blood pressure by activating the natriuretic peptides. CORIN variants have been associated with hypertension and heart disease in African Americans. In this study, we conducted targeted exome sequencing and identified an insertion variant, c.102_103insA, in exon 1 of the CORIN gene. Analysis of two independent cohorts showed that the variant was preferentially present in hypertensive patients (38/795 or 4.78% vs. 4/632 or 0.63% in normal individuals, p = 4.14E-6). The insertion shifted the reading frame, resulting in a corin variant with a truncated cytoplasmic tail. In cell-based studies, the corin variant exhibited poor trafficking in the Golgi, reduced cell surface expression and zymogen activation, and low natriuretic peptide processing activity. Compared with normal individuals with the wild-type allele, individuals with the variant allele had lower levels of plasma corin [0.59 ± 0.07 ng/mL (n = 25) vs. 0.91 ± 0.02 ng/mL (n = 215), p<0.001] and higher levels of plasma N-terminal pro-atrial natriuretic peptide (NT-pro-ANP) [2.39 ± 3.6 nmol/L (n = 21) vs. 0.87 ± 0.6 nmol/L (n = 48), p = 0.005]. These results indicate that the variant altered corin structure and impaired the natriuretic peptide processing activity in vivo. The results highlight corin defects as an important underlying mechanism in hypertension.

Show MeSH
Related in: MedlinePlus