Purification and initial characterization of a novel Porphyromonas gingivalis HmuY protein expressed in Escherichia coli and insect cells.
Porphyromonas gingivalis acquires iron and heme from the host environment using gingipains, lipoproteins, and outer-membrane receptors. Recently, we identified and characterized a heme receptor HmuR. The hmuR gene is localized in an operon together with a hmuY gene encoding a putative heme-binding protein. The aim of this study was to overexpress and perform a preliminary analysis of the recombinant HmuY protein. We constructed and examined several recombinant HmuY variants which were overexpressed and purified from Escherichia coli and insect cells. Recombinant HmuY protein was expressed in insect cells at levels similar to those in E. coli cells. This protein is predominantly present in a monomeric form but also dimerizes and several other oligomerization forms were found. Hemin and ATP binding to the purified HmuY showed that this protein may play a regulatory function in hemin utilization in P. gingivalis.
Olczak
Teresa
T
Laboratory of Biochemistry, Institute of Biochemistry and Molecular Biology, Wroclaw University, Tamka 2, 50-137 Wroclaw, Poland. Teresa.Olczak@bf.uni.wroc.pl
Siudeja
Katarzyna
K
Olczak
Mariusz
M
D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D002352:Carrier Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D002460:Cell Line; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006427:Hemin / Q000737:chemistry / Q000378:metabolism; D007313:Insecta / Q000166:cytology / Q000235:genetics; D016966:Porphyromonas gingivalis / Q000737:chemistry / Q000235:genetics*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins; D011994:Recombinant Proteins; D006427:Hemin; D000255:Adenosine Triphosphate