Streamlining N-terminally anchored yeast surface display via structural insights into S. cerevisiae Pir proteins.
Cell wall
Hsp150
Pir proteins
Saccharomyces cerevisiae
Surface display
Journal
Microbial cell factories
ISSN: 1475-2859
Titre abrégé: Microb Cell Fact
Pays: England
ID NLM: 101139812
Informations de publication
Date de publication:
07 Sep 2023
07 Sep 2023
Historique:
received:
21
07
2023
accepted:
18
08
2023
medline:
11
9
2023
pubmed:
8
9
2023
entrez:
7
9
2023
Statut:
epublish
Résumé
Surface display co-opts yeast's innate ability to embellish its cell wall with mannoproteins, thus converting the yeast's outer surface into a growing and self-sustaining catalyst. However, the efficient toolbox for converting the enzyme of interest into its surface-displayed isoform is currently lacking, especially if the isoform needs to be anchored to the cell wall near the isoform's N-terminus, e.g., through a short GPI-independent protein anchor. Aiming to advance such N-terminally anchored surface display, we employed in silico and machine-learning strategies to study the 3D structure, function, genomic organisation, and evolution of the Pir protein family, whose members evolved to covalently attach themselves near their N-terminus to the β-1,3-glucan of the cell wall. Through the newly-gained insights, we rationally engineered 14 S. cerevisiae Hsp150 (Pir2)-based fusion proteins. We quantified their performance, uncovering guidelines for efficient yeast surface display while developing a construct that promoted a 2.5-fold more efficient display of a reporter protein than the full-length Hsp150. Moreover, we developed a Pir-tag, i.e., a peptide spanning only 4.5 kDa but promoting as efficient surface display of a reporter protein as the full-length Hsp150. These constructs fortify the existing surface display toolbox, allowing for a prompt and routine refitting of intracellular proteins into their N-terminally anchored isoforms.
Identifiants
pubmed: 37679759
doi: 10.1186/s12934-023-02183-2
pii: 10.1186/s12934-023-02183-2
pmc: PMC10483737
doi:
Substances chimiques
Saccharomyces cerevisiae Proteins
0
Glucans
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
174Subventions
Organisme : The Croatian Science Foundation
ID : IP-2019-04-2891
Informations de copyright
© 2023. BioMed Central Ltd., part of Springer Nature.
Références
Nat Methods. 2022 Jun;19(6):679-682
pubmed: 35637307
Appl Microbiol Biotechnol. 2014 Apr;98(7):2897-905
pubmed: 24493571
FEMS Microbiol Rev. 2002 Aug;26(3):239-56
pubmed: 12165426
Antimicrob Agents Chemother. 1972 Apr;1(4):283-8
pubmed: 4208895
Mol Biol Cell. 1998 Dec;9(12):3273-97
pubmed: 9843569
Proc Natl Acad Sci U S A. 2008 Dec 16;105(50):19714-9
pubmed: 19064918
Cold Spring Harb Perspect Biol. 2011 Apr 01;3(4):
pubmed: 21441587
Yeast. 2000 Jun 30;16(9):857-60
pubmed: 10861908
Nat Protoc. 2007;2(1):31-4
pubmed: 17401334
Protein Sci. 2020 Jan;29(1):128-140
pubmed: 31606894
Genetics. 2012 Nov;192(3):775-818
pubmed: 23135325
Proc Natl Acad Sci U S A. 2011 Jan 18;108(3):1058-63
pubmed: 21149693
Science. 2012 Feb 3;335(6068):552-7
pubmed: 22194413
Soft Matter. 2019 Feb 6;15(6):1243-1252
pubmed: 30604826
Yeast. 1993 May;9(5):481-94
pubmed: 8322511
Proc Natl Acad Sci U S A. 2020 Sep 8;117(36):22061-22067
pubmed: 32839341
Nat Ecol Evol. 2022 Apr;6(4):448-460
pubmed: 35210580
Int J Mol Sci. 2020 Nov 26;21(23):
pubmed: 33256216
Methods Mol Biol. 2006;313:333-43
pubmed: 16118442
Mol Microbiol. 1999 Dec;34(5):1049-57
pubmed: 10594829
PLoS One. 2013 Jul 15;8(7):e68734
pubmed: 23869229
J Cell Sci. 2004 Apr 1;117(Pt 9):1665-73
pubmed: 15075228
Bioinformatics. 2018 May 15;34(10):1690-1696
pubmed: 29280997
PLoS One. 2015 Aug 13;10(8):e0135174
pubmed: 26270963
PLoS Genet. 2022 Oct 10;18(10):e1010432
pubmed: 36215302
Nucleic Acids Res. 2011 Jul;39(Web Server issue):W29-37
pubmed: 21593126
Yeast. 1999 Jul;15(10A):813-20
pubmed: 10407261
Genetics. 2003 Jan;163(1):47-54
pubmed: 12586695
Appl Biochem Biotechnol. 2018 Jun;185(2):396-418
pubmed: 29168153
Nat Biotechnol. 2022 Jul;40(7):1023-1025
pubmed: 34980915
Nature. 2004 Apr 8;428(6983):617-24
pubmed: 15004568
Proc Natl Acad Sci U S A. 2012 Aug 14;109(33):13398-403
pubmed: 22847440
J Fungi (Basel). 2021 Feb 09;7(2):
pubmed: 33572482
Bioinformatics. 2018 Nov 15;34(22):3951-3953
pubmed: 29868771
Cell Syst. 2018 Feb 28;6(2):192-205.e3
pubmed: 29361465
Int J Mol Sci. 2021 Nov 01;22(21):
pubmed: 34769302
Nature. 2021 Aug;596(7873):583-589
pubmed: 34265844
Proc Natl Acad Sci U S A. 1992 May 1;89(9):3671-5
pubmed: 1570286
Nucleic Acids Res. 2022 Jan 7;50(D1):D439-D444
pubmed: 34791371
Yeast. 2003 Aug;20(11):973-83
pubmed: 12898712
Biosens Bioelectron. 2022 Oct 15;214:114502
pubmed: 35785751
Front Bioeng Biotechnol. 2022 Jan 10;9:794742
pubmed: 35083204
J Biol Chem. 2006 Apr 28;281(17):11523-9
pubmed: 16495216
J Biol Chem. 1994 May 13;269(19):13887-92
pubmed: 8188666
Nat Microbiol. 2021 Aug;6(8):1055-1065
pubmed: 34326523
Mol Biol Cell. 2021 Mar 1;32(5):363-375
pubmed: 33439673
Adv Mater. 2018 May;30(19):e1704847
pubmed: 29430725
FEMS Yeast Res. 2004 Jan;4(4-5):417-25
pubmed: 14734022
Sci Adv. 2020 Nov 4;6(45):
pubmed: 33148650
Protein Eng Des Sel. 2005 Jul;18(7):337-43
pubmed: 15976011
Biosci Biotechnol Biochem. 2016 Jul;80(7):1243-53
pubmed: 27305282
Curr Opin Struct Biol. 2007 Aug;17(4):467-73
pubmed: 17870469
Comput Biol Chem. 2007 Jun;31(3):151-62
pubmed: 17500039
Mol Biol Cell. 2002 Dec;13(12):4130-40
pubmed: 12475940
Appl Microbiol Biotechnol. 2012 Aug;95(3):577-91
pubmed: 22652839
Appl Environ Microbiol. 1997 Feb;63(2):615-20
pubmed: 9023939
Biomater Res. 2022 Dec 21;26(1):85
pubmed: 36539854
Appl Microbiol Biotechnol. 2001 Nov;57(4):500-5
pubmed: 11762595
Mol Biol Evol. 2021 Jun 25;38(7):3022-3027
pubmed: 33892491
Nat Biotechnol. 1997 Jun;15(6):553-7
pubmed: 9181578
Appl Environ Microbiol. 2002 Sep;68(9):4517-22
pubmed: 12200308
Nat Prod Rep. 2022 Feb 23;39(2):273-310
pubmed: 34755755
Nucleic Acids Res. 2008 Jul 1;36(Web Server issue):W5-9
pubmed: 18440982
Annu Rev Biochem. 1982;51:763-93
pubmed: 7051965
PLoS Comput Biol. 2014 Aug 28;10(8):e1003806
pubmed: 25167361
Nucleic Acids Res. 2014;42(17):10888-902
pubmed: 25190457