Proteases Shape the Chlamydomonas Secretome: Comparison to Classical Neuropeptide Processing Machinery
The recent identification of catalytically active peptidylglycine α-amidating monooxygenase (PAM) in Chlamydomonas reinhardtii, a unicellular green alga, suggested the presence of a PAM-like gene and peptidergic signaling in the last eukaryotic common ancestor (LECA). We identified prototy...
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MDPI AG
2018-09-01
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| Series: | Proteomes |
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| Online Access: | http://www.mdpi.com/2227-7382/6/4/36 |
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| author | Raj Luxmi Crysten Blaby-Haas Dhivya Kumar Navin Rauniyar Stephen M. King Richard E. Mains Betty A. Eipper |
| author_facet | Raj Luxmi Crysten Blaby-Haas Dhivya Kumar Navin Rauniyar Stephen M. King Richard E. Mains Betty A. Eipper |
| author_sort | Raj Luxmi |
| collection | DOAJ |
| description | The recent identification of catalytically active peptidylglycine α-amidating monooxygenase (PAM) in Chlamydomonas reinhardtii, a unicellular green alga, suggested the presence of a PAM-like gene and peptidergic signaling in the last eukaryotic common ancestor (LECA). We identified prototypical neuropeptide precursors and essential peptide processing enzymes (subtilisin-like prohormone convertases and carboxypeptidase B-like enzymes) in the C. reinhardtii genome. Reasoning that sexual reproduction by C. reinhardtii requires extensive communication between cells, we used mass spectrometry to identify proteins recovered from the soluble secretome of mating gametes, and searched for evidence that the putative peptidergic processing enzymes were functional. After fractionation by SDS-PAGE, signal peptide-containing proteins that remained intact, and those that had been subjected to cleavage, were identified. The C. reinhardtii mating secretome contained multiple matrix metalloproteinases, cysteine endopeptidases, and serine carboxypeptidases, along with one subtilisin-like proteinase. Published transcriptomic studies support a role for these proteases in sexual reproduction. Multiple extracellular matrix proteins (ECM) were identified in the secretome. Several pherophorins, ECM glycoproteins homologous to the Volvox sex-inducing pheromone, were present; most contained typical peptide processing sites, and many had been cleaved, generating stable N- or C-terminal fragments. Our data suggest that subtilisin endoproteases and matrix metalloproteinases similar to those important in vertebrate peptidergic and growth factor signaling play an important role in stage transitions during the life cycle of C. reinhardtii. |
| first_indexed | 2024-04-11T13:25:48Z |
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| issn | 2227-7382 |
| language | English |
| last_indexed | 2024-04-11T13:25:48Z |
| publishDate | 2018-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Proteomes |
| spelling | doaj.art-8a728bb8a6624f6c874abe415691b80b2022-12-22T04:22:04ZengMDPI AGProteomes2227-73822018-09-01643610.3390/proteomes6040036proteomes6040036Proteases Shape the Chlamydomonas Secretome: Comparison to Classical Neuropeptide Processing MachineryRaj Luxmi0Crysten Blaby-Haas1Dhivya Kumar2Navin Rauniyar3Stephen M. King4Richard E. Mains5Betty A. Eipper6Department of Neuroscience, University of Connecticut Health Center, Farmington, CT 06030-3401, USADepartment of Biology, Brookhaven National Laboratory, Upton, NY 11973-5000, USADepartment of Molecular Biology and Biophysics, University of Connecticut Health Center, Farmington, CT 06030-3305, USAW.M. Keck Biotechnology Resource Laboratory, Yale University, New Haven, CT 06511-6624, USADepartment of Molecular Biology and Biophysics, University of Connecticut Health Center, Farmington, CT 06030-3305, USADepartment of Neuroscience, University of Connecticut Health Center, Farmington, CT 06030-3401, USADepartment of Neuroscience, University of Connecticut Health Center, Farmington, CT 06030-3401, USAThe recent identification of catalytically active peptidylglycine α-amidating monooxygenase (PAM) in Chlamydomonas reinhardtii, a unicellular green alga, suggested the presence of a PAM-like gene and peptidergic signaling in the last eukaryotic common ancestor (LECA). We identified prototypical neuropeptide precursors and essential peptide processing enzymes (subtilisin-like prohormone convertases and carboxypeptidase B-like enzymes) in the C. reinhardtii genome. Reasoning that sexual reproduction by C. reinhardtii requires extensive communication between cells, we used mass spectrometry to identify proteins recovered from the soluble secretome of mating gametes, and searched for evidence that the putative peptidergic processing enzymes were functional. After fractionation by SDS-PAGE, signal peptide-containing proteins that remained intact, and those that had been subjected to cleavage, were identified. The C. reinhardtii mating secretome contained multiple matrix metalloproteinases, cysteine endopeptidases, and serine carboxypeptidases, along with one subtilisin-like proteinase. Published transcriptomic studies support a role for these proteases in sexual reproduction. Multiple extracellular matrix proteins (ECM) were identified in the secretome. Several pherophorins, ECM glycoproteins homologous to the Volvox sex-inducing pheromone, were present; most contained typical peptide processing sites, and many had been cleaved, generating stable N- or C-terminal fragments. Our data suggest that subtilisin endoproteases and matrix metalloproteinases similar to those important in vertebrate peptidergic and growth factor signaling play an important role in stage transitions during the life cycle of C. reinhardtii.http://www.mdpi.com/2227-7382/6/4/36peptidylglycine α-amidating monooxygenaseciliamatingsignal peptideprohormone convertasecarboxypeptidasematrix metalloproteinasesubtilisinpherophorin |
| spellingShingle | Raj Luxmi Crysten Blaby-Haas Dhivya Kumar Navin Rauniyar Stephen M. King Richard E. Mains Betty A. Eipper Proteases Shape the Chlamydomonas Secretome: Comparison to Classical Neuropeptide Processing Machinery Proteomes peptidylglycine α-amidating monooxygenase cilia mating signal peptide prohormone convertase carboxypeptidase matrix metalloproteinase subtilisin pherophorin |
| title | Proteases Shape the Chlamydomonas Secretome: Comparison to Classical Neuropeptide Processing Machinery |
| title_full | Proteases Shape the Chlamydomonas Secretome: Comparison to Classical Neuropeptide Processing Machinery |
| title_fullStr | Proteases Shape the Chlamydomonas Secretome: Comparison to Classical Neuropeptide Processing Machinery |
| title_full_unstemmed | Proteases Shape the Chlamydomonas Secretome: Comparison to Classical Neuropeptide Processing Machinery |
| title_short | Proteases Shape the Chlamydomonas Secretome: Comparison to Classical Neuropeptide Processing Machinery |
| title_sort | proteases shape the chlamydomonas secretome comparison to classical neuropeptide processing machinery |
| topic | peptidylglycine α-amidating monooxygenase cilia mating signal peptide prohormone convertase carboxypeptidase matrix metalloproteinase subtilisin pherophorin |
| url | http://www.mdpi.com/2227-7382/6/4/36 |
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