How many protein molecules are secreted by single Helicobacter pylori cells: Quantification of serine protease HtrA

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Details zur Publikation

Autorinnen und Autoren: Neddermann M, Backert S
Zeitschrift: Cellular Microbiology
Jahr der Veröffentlichung: 2019
Band: 21
Heftnummer: 7
ISSN: 1462-5814
eISSN: 1462-5822


Abstract

Infection with Helicobacter pylori represents a major risk for developing peptic ulcer disease, gastric adenocarcinoma, and various other gastric and nongastric sicknesses. A series of H. pylori virulence factors can be secreted into the cell culture supernatant, and the secretome contains more than 100 different proteins. However, the quantities of proteins secreted by the bacteria over time are unknown. One of these factors is the serine protease high-temperature requirement A (HtrA), encoded by an essential bifunctional gene with crucial intracellular and extracellular activities. We have demonstrated recently that secreted HtrA can cleave off the ectodomains of the tight junction proteins occludin and claudin-8, as well as of the tumour suppressor and adherens junction protein E-cadherin on polarised gastric epithelial cells. The exact mechanism of secretion and the quantity of secreted HtrA, however, have not been studied in detail. Here, we applied protein purification and quantitative Western blotting to determine the number of HtrA molecules secreted by H. pylori cells in liquid culture during a time course. Over a period of 8 hr, actively dividing bacteria secreted HtrA at a similar rate, on average about 9,600 HtrA molecules per cell. We determined minor variation over time corresponding to 9,931 ± 1,768 at an OD600 of 0.4 after 2 hr, 9,403 ± 2,356 2 hr later, and 9,644 ± 2,067 molecules per cell after 8 hr of culturing, when the culture had reached an OD600 of 0.8. This is the first report on the quantification of a secreted virulence protein from the important gastric pathogen H. pylori. Because HtrA has been considered as a promising new target for antibacterial therapy, knowledge about secreted protein quantities is crucial for optimising corresponding treatment regimes.


FAU-Autorinnen und Autoren / FAU-Herausgeberinnen und Herausgeber

Backert, Steffen Prof. Dr.
Lehrstuhl für Mikrobiologie
Neddermann, Matthias
Lehrstuhl für Mikrobiologie


Zitierweisen

APA:
Neddermann, M., & Backert, S. (2019). How many protein molecules are secreted by single Helicobacter pylori cells: Quantification of serine protease HtrA. Cellular Microbiology, 21(7). https://dx.doi.org/10.1111/cmi.13022

MLA:
Neddermann, Matthias, and Steffen Backert. "How many protein molecules are secreted by single Helicobacter pylori cells: Quantification of serine protease HtrA." Cellular Microbiology 21.7 (2019).

BibTeX: 

Zuletzt aktualisiert 2019-21-06 um 09:03