Structure-function analysis of PrsA reveals roles for the parvulin-like and flanking N- and C-terminal domains in protein folding and secretion in Bacillus subtilis

Marika Vitikainen, Ilkka Lappalainen, Raili Seppala, Haike Antelmann, Harry Boer, Suvi Taira, Harri Savilahti, Michael Hecker, Mauno Vihinen, Matti Sarvas, Vesa P. Kontinen (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

74 Citations (Scopus)

Abstract

The PrsA protein of Bacillus subtilis is an essential membrane-bound lipoprotein that is assumed to assist post-translocational folding of exported proteins and stabilize them in the compartment between the cytoplasmic membrane and cell wall. This folding activity is consistent with the homology of a segment of PrsA with parvulin-type peptidyl-prolyl cis/trans isomerases (PPIase). In this study, molecular modeling showed that the parvulin-like region can adopt a parvulin-type fold with structurally conserved active site residues. PrsA exhibits PPIase activity in a manner dependent on the parvulin-like domain. We constructed deletion, peptide insertion, and amino acid substitution mutations and demonstrated that the parvulin-like domain as well as flanking N- and C-terminal domains are essential for in vivo PrsA function in protein secretion and growth. Surprisingly, none of the predicted active site residues of the parvulin-like domain was essential for growth and protein secretion, although several active site mutations reduced or abolished the PPIase activity or the ability of PrsA to catalyze proline-limited protein folding in vitro. Our results indicate that PrsA is a PPIase, but the essential role in vivo seems to depend on some non-PPIase activity of both the parvulin-like and flanking domains.
Original languageEnglish
Pages (from-to)19302 - 19314
Number of pages13
JournalJournal of Biological Chemistry
Volume279
Issue number18
DOIs
Publication statusPublished - 2004
MoE publication typeA1 Journal article-refereed

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Peptidylprolyl Isomerase
Protein folding
Protein Folding
Bacilli
Bacillus subtilis
Catalytic Domain
cis-trans-Isomerases
Proteins
Membranes
Molecular modeling
Mutation
Proline
Lipoproteins
Amino Acid Substitution
Growth
Substitution reactions
Cell Wall
Cells
Amino Acids
Peptides

Keywords

  • protein secretion
  • proteins
  • Bacillus subtilis

Cite this

Vitikainen, Marika ; Lappalainen, Ilkka ; Seppala, Raili ; Antelmann, Haike ; Boer, Harry ; Taira, Suvi ; Savilahti, Harri ; Hecker, Michael ; Vihinen, Mauno ; Sarvas, Matti ; Kontinen, Vesa P. / Structure-function analysis of PrsA reveals roles for the parvulin-like and flanking N- and C-terminal domains in protein folding and secretion in Bacillus subtilis. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 18. pp. 19302 - 19314.
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abstract = "The PrsA protein of Bacillus subtilis is an essential membrane-bound lipoprotein that is assumed to assist post-translocational folding of exported proteins and stabilize them in the compartment between the cytoplasmic membrane and cell wall. This folding activity is consistent with the homology of a segment of PrsA with parvulin-type peptidyl-prolyl cis/trans isomerases (PPIase). In this study, molecular modeling showed that the parvulin-like region can adopt a parvulin-type fold with structurally conserved active site residues. PrsA exhibits PPIase activity in a manner dependent on the parvulin-like domain. We constructed deletion, peptide insertion, and amino acid substitution mutations and demonstrated that the parvulin-like domain as well as flanking N- and C-terminal domains are essential for in vivo PrsA function in protein secretion and growth. Surprisingly, none of the predicted active site residues of the parvulin-like domain was essential for growth and protein secretion, although several active site mutations reduced or abolished the PPIase activity or the ability of PrsA to catalyze proline-limited protein folding in vitro. Our results indicate that PrsA is a PPIase, but the essential role in vivo seems to depend on some non-PPIase activity of both the parvulin-like and flanking domains.",
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Structure-function analysis of PrsA reveals roles for the parvulin-like and flanking N- and C-terminal domains in protein folding and secretion in Bacillus subtilis. / Vitikainen, Marika; Lappalainen, Ilkka; Seppala, Raili; Antelmann, Haike; Boer, Harry; Taira, Suvi; Savilahti, Harri; Hecker, Michael; Vihinen, Mauno; Sarvas, Matti; Kontinen, Vesa P. (Corresponding Author).

In: Journal of Biological Chemistry, Vol. 279, No. 18, 2004, p. 19302 - 19314.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Structure-function analysis of PrsA reveals roles for the parvulin-like and flanking N- and C-terminal domains in protein folding and secretion in Bacillus subtilis

AU - Vitikainen, Marika

AU - Lappalainen, Ilkka

AU - Seppala, Raili

AU - Antelmann, Haike

AU - Boer, Harry

AU - Taira, Suvi

AU - Savilahti, Harri

AU - Hecker, Michael

AU - Vihinen, Mauno

AU - Sarvas, Matti

AU - Kontinen, Vesa P.

PY - 2004

Y1 - 2004

N2 - The PrsA protein of Bacillus subtilis is an essential membrane-bound lipoprotein that is assumed to assist post-translocational folding of exported proteins and stabilize them in the compartment between the cytoplasmic membrane and cell wall. This folding activity is consistent with the homology of a segment of PrsA with parvulin-type peptidyl-prolyl cis/trans isomerases (PPIase). In this study, molecular modeling showed that the parvulin-like region can adopt a parvulin-type fold with structurally conserved active site residues. PrsA exhibits PPIase activity in a manner dependent on the parvulin-like domain. We constructed deletion, peptide insertion, and amino acid substitution mutations and demonstrated that the parvulin-like domain as well as flanking N- and C-terminal domains are essential for in vivo PrsA function in protein secretion and growth. Surprisingly, none of the predicted active site residues of the parvulin-like domain was essential for growth and protein secretion, although several active site mutations reduced or abolished the PPIase activity or the ability of PrsA to catalyze proline-limited protein folding in vitro. Our results indicate that PrsA is a PPIase, but the essential role in vivo seems to depend on some non-PPIase activity of both the parvulin-like and flanking domains.

AB - The PrsA protein of Bacillus subtilis is an essential membrane-bound lipoprotein that is assumed to assist post-translocational folding of exported proteins and stabilize them in the compartment between the cytoplasmic membrane and cell wall. This folding activity is consistent with the homology of a segment of PrsA with parvulin-type peptidyl-prolyl cis/trans isomerases (PPIase). In this study, molecular modeling showed that the parvulin-like region can adopt a parvulin-type fold with structurally conserved active site residues. PrsA exhibits PPIase activity in a manner dependent on the parvulin-like domain. We constructed deletion, peptide insertion, and amino acid substitution mutations and demonstrated that the parvulin-like domain as well as flanking N- and C-terminal domains are essential for in vivo PrsA function in protein secretion and growth. Surprisingly, none of the predicted active site residues of the parvulin-like domain was essential for growth and protein secretion, although several active site mutations reduced or abolished the PPIase activity or the ability of PrsA to catalyze proline-limited protein folding in vitro. Our results indicate that PrsA is a PPIase, but the essential role in vivo seems to depend on some non-PPIase activity of both the parvulin-like and flanking domains.

KW - protein secretion

KW - proteins

KW - Bacillus subtilis

U2 - 10.1074/jbc.M400861200

DO - 10.1074/jbc.M400861200

M3 - Article

VL - 279

SP - 19302

EP - 19314

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 18

ER -