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Analysis of the polycystin complex (PCC) in human urinary exosome–like vesicles (ELVs)

The polycystin–1 (PC1), polycystin–2 (PC2) and fibrocystin proteins, the respective products of the PKD1, PKD2 and PKHD1 genes, are abundant in urinary exosome–like vesicles (ELVs) where they form the polycystin complex (PCC). ELVs are 100 nm diameter membrane vesicles shed into the urine by the cel...

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Autores principales: Lea, Wendy A., McGreal, Kerri, Sharma, Madhulika, Parnell, Stephen C., Zelenchuk, Lesya, Charlesworth, M. Cristine, Madden, Benjamin J., Johnson, Kenneth L., McCormick, Daniel J., Hogan, Marie C., Ward, Christopher J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6992733/
https://www.ncbi.nlm.nih.gov/pubmed/32001768
http://dx.doi.org/10.1038/s41598-020-58087-3
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author Lea, Wendy A.
McGreal, Kerri
Sharma, Madhulika
Parnell, Stephen C.
Zelenchuk, Lesya
Charlesworth, M. Cristine
Madden, Benjamin J.
Johnson, Kenneth L.
McCormick, Daniel J.
Hogan, Marie C.
Ward, Christopher J.
author_facet Lea, Wendy A.
McGreal, Kerri
Sharma, Madhulika
Parnell, Stephen C.
Zelenchuk, Lesya
Charlesworth, M. Cristine
Madden, Benjamin J.
Johnson, Kenneth L.
McCormick, Daniel J.
Hogan, Marie C.
Ward, Christopher J.
author_sort Lea, Wendy A.
collection PubMed
description The polycystin–1 (PC1), polycystin–2 (PC2) and fibrocystin proteins, the respective products of the PKD1, PKD2 and PKHD1 genes, are abundant in urinary exosome–like vesicles (ELVs) where they form the polycystin complex (PCC). ELVs are 100 nm diameter membrane vesicles shed into the urine by the cells lining the nephron. Using MS/MS analysis of ELVs from individuals with PKD1 mutations and controls, we show that in addition to the well-described GPS/GAIN cleavage event in PC1 at 3048 aa and the proprotein convertase cleavage (PPC) event in fibrocystin at 3616 aa, there are multiple other cleavage events in these proteins. The C–terminal 11 transmembrane portion of PC1 undergoes three cleavage events in vivo. The absence of peptides from the C–terminal cytoplasmic tail of fibrocystin implies a cleavage event close to its single TM domain prior to loading onto the ELVs. There is also evidence that the C–terminal tail of PC2 is also cleaved in ELVs. Native gel analysis of the PCC shows that the entire complex is  > 2 MDa in size and that N–terminal GPS/GAIN cleaved PC1 and PPC cleaved fibrocystin ectodomains can be released under non-reducing conditions and resolve at 300 kDa. This paper shows that the three major human cystogene proteins are detectable in human urinary ELVs and that all three undergo post-translational proteolytic processing. Human urinary ELVs may be a useful source of material in the search for proteins that interact with the PCC.
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spelling pubmed-69927332020-02-05 Analysis of the polycystin complex (PCC) in human urinary exosome–like vesicles (ELVs) Lea, Wendy A. McGreal, Kerri Sharma, Madhulika Parnell, Stephen C. Zelenchuk, Lesya Charlesworth, M. Cristine Madden, Benjamin J. Johnson, Kenneth L. McCormick, Daniel J. Hogan, Marie C. Ward, Christopher J. Sci Rep Article The polycystin–1 (PC1), polycystin–2 (PC2) and fibrocystin proteins, the respective products of the PKD1, PKD2 and PKHD1 genes, are abundant in urinary exosome–like vesicles (ELVs) where they form the polycystin complex (PCC). ELVs are 100 nm diameter membrane vesicles shed into the urine by the cells lining the nephron. Using MS/MS analysis of ELVs from individuals with PKD1 mutations and controls, we show that in addition to the well-described GPS/GAIN cleavage event in PC1 at 3048 aa and the proprotein convertase cleavage (PPC) event in fibrocystin at 3616 aa, there are multiple other cleavage events in these proteins. The C–terminal 11 transmembrane portion of PC1 undergoes three cleavage events in vivo. The absence of peptides from the C–terminal cytoplasmic tail of fibrocystin implies a cleavage event close to its single TM domain prior to loading onto the ELVs. There is also evidence that the C–terminal tail of PC2 is also cleaved in ELVs. Native gel analysis of the PCC shows that the entire complex is  > 2 MDa in size and that N–terminal GPS/GAIN cleaved PC1 and PPC cleaved fibrocystin ectodomains can be released under non-reducing conditions and resolve at 300 kDa. This paper shows that the three major human cystogene proteins are detectable in human urinary ELVs and that all three undergo post-translational proteolytic processing. Human urinary ELVs may be a useful source of material in the search for proteins that interact with the PCC. Nature Publishing Group UK 2020-01-30 /pmc/articles/PMC6992733/ /pubmed/32001768 http://dx.doi.org/10.1038/s41598-020-58087-3 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Lea, Wendy A.
McGreal, Kerri
Sharma, Madhulika
Parnell, Stephen C.
Zelenchuk, Lesya
Charlesworth, M. Cristine
Madden, Benjamin J.
Johnson, Kenneth L.
McCormick, Daniel J.
Hogan, Marie C.
Ward, Christopher J.
Analysis of the polycystin complex (PCC) in human urinary exosome–like vesicles (ELVs)
title Analysis of the polycystin complex (PCC) in human urinary exosome–like vesicles (ELVs)
title_full Analysis of the polycystin complex (PCC) in human urinary exosome–like vesicles (ELVs)
title_fullStr Analysis of the polycystin complex (PCC) in human urinary exosome–like vesicles (ELVs)
title_full_unstemmed Analysis of the polycystin complex (PCC) in human urinary exosome–like vesicles (ELVs)
title_short Analysis of the polycystin complex (PCC) in human urinary exosome–like vesicles (ELVs)
title_sort analysis of the polycystin complex (pcc) in human urinary exosome–like vesicles (elvs)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6992733/
https://www.ncbi.nlm.nih.gov/pubmed/32001768
http://dx.doi.org/10.1038/s41598-020-58087-3
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