PXD032195

PXD032195 is an original dataset announced via ProteomeXchange.

Title	UNASSAMBLED CELL WALL PROTEINS FORM AGGREGATES IN THE EXTRACELLULAR SPACE OF CHLAMYDOMONAS REINHARDTII STRAIN UVM4
Description	The green microalga Chlamydomonas reinhardtii is emerging as a promising cell biofactory for secreted recombinant protein (RP) production. In recent years, the generation of the broadly-used cell wall-deficient mutant strain UVM4 has allowed for a drastic increase in secreted RP yields. However, purification of secreted RPs from the extracellular space of C. reinhardtii strain UVM4 is challenging. Previous studies suggest that secreted RPs are trapped in a matrix of cell wall protein aggregates populating the secretome of strain UVM4, making it difficult to isolate and purify the RPs. To better understand the nature and behaviour of these extracellular protein aggregates, we analysed and compared the extracellular proteome of the strain UVM4 to its cell-walled ancestor, C. reinhardtii strain 137c. When grown under the same conditions, strain UVM4 produced a unique extracellular proteomic profile, including a higher abundance of secreted cell wall glycoproteins. Further characterization of high molecular weight extracellular protein aggregates in strain UVM4 revealed that they are largely comprised of pherophorins, a specific class of cell wall glycoproteins. Our results offer important new insights into the extracellular space of strain UVM4, including strain-specific secreted cell wall proteins and the composition of the aggregates possibly related to impaired RP purification. The discovery of pherophorins as a major component of extracellular protein aggregates will inform future strategies to remove or prevent aggregate formation, enhance purification of secreted RPs, and improve yields of recombinant biopharmaceuticals in this emerging cell biofactory.
HostingRepository	PRIDE
AnnounceDate	2022-08-11
AnnouncementXML	Submission_2022-08-11_06:10:49.961.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD032195
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Matthew Padula
SpeciesList	scientific name: Chlamydomonas reinhardtii; NCBI TaxID: 3055;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue; deamidated residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2022-03-11 08:53:44	ID requested
⏵ 1	2022-08-11 06:10:50	announced

Barolo L, Commault AS, Abbriano RM, Padula MP, Kim M, Kuzhiumparambil U, Ralph PJ, Pernice M, Unassembled cell wall proteins form aggregates in the extracellular space of Chlamydomonas reinhardtii strain UVM4. Appl Microbiol Biotechnol, 106(11):4145-4156(2022) [pubmed]

submitter keyword: Microalgae, recombinant protein, secretome, Chlamydomonas, glycoproteins, proteomics, pherophorins

Peter J. Ralph
contact affiliation	Climate Change Cluster, University of Technology Sydney, 15 Broadway, Ultimo 2007, Sydney, NSW, Australia
contact email	peter.ralph@uts.edu.au
lab head
Matthew Padula
contact affiliation	University of technology Sydney
contact email	matthew.padula@uts.edu.au
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD032195
     1. Label: PRIDE project
     2. Name: UNASSAMBLED CELL WALL PROTEINS FORM AGGREGATES IN THE EXTRACELLULAR SPACE OF CHLAMYDOMONAS REINHARDTII STRAIN UVM4