Secretome of Transmissible <i>Pseudomonas aeruginosa</i> AES-1R Grown in a Cystic Fibrosis
Lung-Like Environment
Nichollas
E. Scott
Nathan
J. Hare
Melanie Y. White
Jim Manos
Stuart J. Cordwell
10.1021/pr4007365.s001
https://acs.figshare.com/articles/journal_contribution/Secretome_of_Transmissible_i_Pseudomonas_aeruginosa_i_AES_1R_Grown_in_a_Cystic_Fibrosis_Lung_Like_Environment/2346649
<i>Pseudomonas aeruginosa</i> is the predominant cause
of mortality in patients with cystic fibrosis (CF). We examined the
secretome of an acute, transmissible CF <i>P. aeruginosa</i> (Australian epidemic strain 1-R; AES-1R) compared with laboratory-adapted
PAO1. Culture supernatant proteins from rich (LB) and minimal (M9)
media were compared using 2-DE and 2DLC-MS/MS, which revealed elevated
abundance of PasP protease and absence of AprA protease in AES-1R.
CF lung-like artificial sputum medium (ASMDM) contains serum and mucin
that generally preclude proteomics of secreted proteins. ASMDM culture
supernatants were subjected to 2DLC-MS/MS, which allowed the identification
of 57 <i>P. aeruginosa</i> proteins, and qualitative spectral
counting was used to estimate relative abundance. AES-1R-specific
AES_7139 and PasP were more abundant in AES-1R ASMDM culture supernatants,
while AprA could only be identified in PAO1. Relative quantitation
was performed using selected reaction monitoring. Significantly elevated
levels of PasP, LasB, chitin-binding protein (CbpD), and PA4495 were
identified in AES-1R ASMDM supernatants. Quantitative PCR showed elevated <i>pasP</i> in AES-1R during early (18 h) ASMDM growth, while no
evidence of <i>aprA</i> expression could be observed. Genomic
screening of CF isolates revealed <i>aes_7139</i> was present
in all AES-1 and one pair of sequential nonepidemic isolates. Secreted
proteins may be crucial in aiding CF-associated <i>P. aeruginosa</i> to establish infection and for adaptation to the CF lung.
2013-12-06 00:00:00
Secreted proteins
AES
ASMDM growth
reaction monitoring
CF lung
57 P
sequential nonepidemic
sputum medium
aprA expression
AprA protease
PasP protease
Quantitative PCR
aeruginosa proteins
ASMDM culture supernatants
DE
LB
PAO 1.
CF P
PA 4495
Genomic screening