Molecular characterization of a heme-binding protein of Bacteroides fragilis BE1.
An iron-repressible 44-kDa outer membrane protein plays a crucial role in the acquisition of heme by the anaerobic bacterium Bacteroides fragilis. The DNA sequence of the gene encoding the 44-kDa protein (hupA) was determined. The hupA gene encodes a protein of 431 amino acid residues with a calculated molecular mass of 48,189 Da. The hupA gene is preceded by an open reading frame of 480 bp that probably encodes a protein with a calculated molecular mass of 18,073 Da. hupA and this open reading frame are likely organized in an operon, and a sequence homologous to the Escherichia coli consensus Fur box was present in the putative promoter region of the operon. Heme-binding studies showed that HupA binds heme. Analysis of the deduced amino acid sequence revealed signature heme-binding consensus motifs, characteristic of heme lyases. Subcellular localization studies in E. coli revealed that HupA was mainly found in the cytoplasmic membrane but not in the outer membrane of E. coli. This suggested that B. fragilis uses another strategy for the translocation of this outer membrane protein across its cell envelope than E. coli does. HupA did not have significant homology with other putative bacterial heme receptors.
Otto
B R
BR
Department of Molecular Microbiology, Institute of Molecular Biological Sciences, Biocentrum Amsterdam, Vrije Universiteit, The Netherlands. brotto@bio.vu.nl
Kusters
J G
JG
Luirink
J
J
de Graaf
F K
FK
Oudega
B
B
D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D001441:Bacteroides fragilis / Q000737:chemistry*; D001483:Base Sequence; D002352:Carrier Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D000081062:Heme-Binding Proteins; D006420:Hemeproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight
D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D000081062:Heme-Binding Proteins; D006420:Hemeproteins; C099188:HupA protein, bacteria; C490937:hupA protein, E coli