Abstract

Iron is an important nutrient required by all forms of life.In the case of human hosts,the free iron availability is 10(-18) M,which is far less than what is needed for the survival of the invading bacterial pathogen. To survive in such conditions, bacteria express new proteins in their outer membrane and also secrete iron chelators called siderophores. RESULTS/ DISCUSSION: Acinetobacter baumannii ATCC 19606, a nosocomial pathogen which grows under iron restricted conditions, expresses four new outer membrane proteins,with molecular weight ranging from 77 kDa to 88 kDa, that are called Iron Regulated Outer Membrane Proteins (IROMPs). We studied the functional and immunological properties of IROMPs expressed by A.baumanii ATCC 19606. The bands corresponding to IROMPs were eluted from SDS-PAGE and were used to immunize BALB/c mice for the production of monoclonal antibodies. Hybridomas secreting specific antibodies against these IROMPs were selected after screening by ELISA and their reactivity was confirmed by Western Blot. The antibodies then generated belonged to IgM isotype and showed bactericidical and opsonising activities against A.baumanii in vitro. These antibodies also blocked siderophore mediated iron uptake via IROMPs in bacteria. This proves that iron uptake via IROMPs,which is mediated through siderophores,may have an important role in the survival of A.baumanii inside the host,and helps establishing the infection.

Highlights

  • Iron is one of the essential nutrients required by the bacteria to be able to multiply and invade a vertebrate host

  • The role of Iron Regulated Outer Membrane Proteins (IROMPs) in iron uptake have been reported for other bacteria like Escherichia coli, Pseudomonas aeruginosa and Neisse

  • Expression of IROMPs and effect of Iron on growth OMP profiles of A. baumannii grown in CDM-Fe and CDM+Fe were compared on SDS-PAGE and it was seen that in absence of iron, bacteria developed 4 new outer membrane proteins which were absent in iron replete conditions

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Summary

Introduction

Iron is one of the essential nutrients required by the bacteria to be able to multiply and invade a vertebrate host. One of the methods of achieving this, is by secretion of low molecular weight high affinity iron chelators termed siderophores and their specific cell surface receptor, Iron Regulated Outer Membrane Proteins (IROMPs). These receptors are expressed under iron restricted conditions [6,7,8,9]. Iron is an important nutrient required by all forms of life.In the case of human hosts,the free iron availability is 10-18M,which is far less than what is needed for the survival of the invading bacterial pathogen.To survive in such conditions, bacteria express new proteins in their outer membrane and secrete iron chelators called siderophores

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