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Title: Proteases Shape the Chlamydomonas Secretome: Comparison to Classical Neuropeptide Processing Machinery

Abstract

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 vertebratemore » peptidergic and growth factor signaling play an important role in stage transitions during the life cycle of C. reinhardtii.« less

Authors:
 [1];  [2];  [1];  [3];  [1]; ORCiD logo [1];  [1]
  1. Univ. of Connecticut Health Center, Farmington, CT (United States)
  2. Brookhaven National Lab. (BNL), Upton, NY (United States)
  3. Yale Univ., New Haven, CT (United States)
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1473636
Report Number(s):
BNL-209077-2018-JAAM
Journal ID: ISSN 2227-7382; PROTHC
Grant/Contract Number:  
SC0012704
Resource Type:
Accepted Manuscript
Journal Name:
Proteomes
Additional Journal Information:
Journal Volume: 6; Journal Issue: 4; Journal ID: ISSN 2227-7382
Publisher:
MDPI
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; peptidylglycine; amidating monooxygenase; cilia; mating; signal peptide; prohormone; convertase; carboxypeptidase; matrix metalloproteinase; subtilisin; pherophorin

Citation Formats

Luxmi, Raj, Blaby-Haas, Crysten, Kumar, Dhivya, Rauniyar, Navin, King, Stephen M., Mains, Richard E., and Eipper, Betty A. Proteases Shape the Chlamydomonas Secretome: Comparison to Classical Neuropeptide Processing Machinery. United States: N. p., 2018. Web. doi:10.3390/proteomes6040036.
Luxmi, Raj, Blaby-Haas, Crysten, Kumar, Dhivya, Rauniyar, Navin, King, Stephen M., Mains, Richard E., & Eipper, Betty A. Proteases Shape the Chlamydomonas Secretome: Comparison to Classical Neuropeptide Processing Machinery. United States. doi:10.3390/proteomes6040036.
Luxmi, Raj, Blaby-Haas, Crysten, Kumar, Dhivya, Rauniyar, Navin, King, Stephen M., Mains, Richard E., and Eipper, Betty A. Sun . "Proteases Shape the Chlamydomonas Secretome: Comparison to Classical Neuropeptide Processing Machinery". United States. doi:10.3390/proteomes6040036. https://www.osti.gov/servlets/purl/1473636.
@article{osti_1473636,
title = {Proteases Shape the Chlamydomonas Secretome: Comparison to Classical Neuropeptide Processing Machinery},
author = {Luxmi, Raj and Blaby-Haas, Crysten and Kumar, Dhivya and Rauniyar, Navin and King, Stephen M. and Mains, Richard E. and Eipper, Betty A.},
abstractNote = {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.},
doi = {10.3390/proteomes6040036},
journal = {Proteomes},
number = 4,
volume = 6,
place = {United States},
year = {2018},
month = {9}
}

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