Lysine-specific gingipain K and heme/hemoglobin receptor HmuR are involved in heme utilization in Porphyromonas gingivalis.

• Authors: Waltena Simpson , Teresa Olczak , Caroline A Genco
• Languages of publication: EN

Abstracts

EN

We have previously reported on the identification and characterization of the Porphyromonas gingivalis A7436 strain outer membrane receptor HmuR, which is involved in the acquisition of hemin and hemoglobin. We demonstrated that HmuR interacts with the lysine- (Kgp) and arginine- (HRgpA) specific proteases (gingipains) and that Kgp and HRgpA can bind and degrade hemoglobin. Here, we report on the physiological significance of the HmuR-Kgp complex in heme utilization in P. gingivalis through the construction and characterization of a defined kgp mutant and a hmuR kgp double mutant in P. gingivalis A7436. The P. gingivalis kgp mutant exhibited a decreased ability to bind both hemin and hemoglobin. Growth of this strain with hemoglobin was delayed and its ability to utilize hemin as a sole iron source was diminished as compared to the wild type strain. Inactivation of both the hmuR and kgp genes resulted in further decreased ability of P. gingivalis to bind hemoglobin and hemin, as well as diminished ability to utilize either hemin or hemoglobin as a sole iron source. Collectively, these in vivo results further confirmed that both HmuR and Kgp are involved in the utilization of hemin and hemoglobin in P. gingivalis A7436.

Keywords

EN

Kgp; heme; Porphyromonas gingivalis; HmuR; gingipain

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2004
• Volume: 51
• Issue: 1
• Pages: 253-262

Dates

published

2004

received

2003-10-17

revised

2003-12-05

accepted

2004-01-10

Contributors

author

Waltena Simpson

• Department of Medicine, Section of Infectious Diseases, Boston University School of Medicine, Boston, MA 02118, U.S.A.

author

Teresa Olczak

• Department of Medicine, Section of Infectious Diseases, Boston University School of Medicine, Boston, MA 02118, U.S.A.

author

Caroline A Genco

• Department of Medicine, Section of Infectious Diseases, Boston University School of Medicine, Boston, MA 02118, U.S.A.

References

• Aduse-Opoku J, Slaney J, Rangarajan M, Muir J, Young K, Curtis MA. (1997) The Tla protein of Porphyromonas gingivalis W50: a homolog of the R1 protease precursor (PrpR1) is an outer membrane receptor required for growth on low levels of hemin. J Bacteriol.; 179: 4778-88.
• Aduse-Opoku J, Davies NN, Gallagher A, Hashim A, Evans HE, Rangarajan M, Slaney JM, Curtis MA. (2000) Generation of Lys-gingipain protease activity in Porphyromonas gingivalis W50 independent of Arg-gingipain protease activities. Microbiology.; 146: 1933-40.
• Brochu V, Grenier D, Nakayama K, Mayrand D. (2001) Acquisition of iron from human transferrin by Porphyromonas gingivalis: a role for Arg- and Lys-gingipain activities. Oral Microbiol Immunol.; 16: 79-87.
• Dashper SG, Hendtlass A, Slakeski N, Jackson C, Cross KJ, Brownfield L, Hamilton R, Barr I, Reynolds EC. (2000) Characterization of a novel outer membrane hemin binding protein of Porphyromonas gingivalis. J Bacteriol.; 182: 6456-62.
• DeCarlo AA, Paramaesvaran M, Yun PLW, Collyer C, Hunter N. (1999) Porphyrin-mediated binding to hemoglobin by theHA2 domain of cysteine proteinases (gingipains) and hemagglutinins from the periodontal pathogen Porphyromonas gingivalis. J Bacteriol.; 181: 3784-91.
• Genco CA, Simpson W, Forng RY, Egal M, Odusanya M. (1995) Characterization of a Tn4351-generated hemin uptake mutant of Porphyromonas gingivalis: Evidence for the coordinate regulation of virulence factors by hemin. Infect Immun.; 63: 2459-66.
• Ghigo MJ, Letoffe S, Wandersman C. (1997) A new type of hemophore-dependent heme acquisition system of Serratia marcescens reconstituted in Escherichia coli. J Bacteriol.; 179: 3572-9.
• Han N, Whitlock J, Progulske-Fox A. (1996) The hemagglutinin gene A (hagA) of Porphyromonas gingivalis 381 contains four large, contiguous, direct repeats. Infect Immun.; 64: 4000-7.
• Hanley SA, Aduse-Opoku J, Curtis M. (1999) A 55-kilodalton immunodominant antigen of Porphyromonas gingivalis W50 has arisen via horizontal gene transfer. Infect Immun.; 67: 1157-71.
• Karunakaran T, Madden T, Kuramitsu K. (1997) Isolation and characterization of a hemin-regulated gene, hemR, from Porphyromonas gingivalis. J Bacteriol.; 179: 1898-908.
• Kuboniwa M, Amano A, Shizukuishi S. (1998) Hemoglobin-binding protein purified from Porphyromonas gingivalis is identical to lysine-specific cysteine proteinase (Lys- gingipain). Biochem Biophys Res Commun.; 249: 38-43.
• Letoffe S, Ghigo JM, Wandersman C. (1994) Iron acquisition from heme and hemoglobin by a Serratia marcescens extracellular protein. Proc Natl Acad Sci U S A.; 91: 9876-80.
• Letoffe S, Nato F, Goldberg ME, Wandersman C. (1999) Interactions of HasA, a bacterial hemophore, with hemoglobin and with its outer membrane receptor HasR. Mol Microbiol.; 33: 546-55.
• Lewis LA, Dyer DW. (1995) Identification of an iron-regulated outer membrane protein of Neisseria meningitidis involved in the utilization of hemoglobin complexed to haptoglobin. J Bacteriol.; 177: 1299-306.
• Lewis LA, Gray E, Wang YP, Roe BA, Dyer DW. (1997) Molecular characterization of hpuAB, the haemoglobin-haptoglobin utilization operon of Neisseria meningitidis. Mol Microbiol.; 23: 737-49.
• Lewis JP, Dawson JA, Hannis JC, Muddiman D, Macrina FL. (1999) Hemoglobinase activity of the lysine gingipain protease (Kgp) of Porphyromonas gingivalis. J Bacteriol.; 181: 4905-13.
• Morton DJ, Whitby PW, Jin H, Ren Z, Stull TL. (1999) Effect of multiple mutations in the hemoglobin and hemoglobin-haptoglobin binding proteins, HgpA, HgpB, and HgpC, of Haemophilus influenzae type b. Infect Immun.; 67: 2729-39.
• Nakayama K, Ratnayake DB, Tsukuba T, Kadowaki T, Yamamoto K, Fujimura S. (1998) Hemoglobin receptor protein is intragenically encoded by the cysteine proteinase encoding genes and the haemagglutinin-encoding gene of Porphyromonas gingivalis. Mol Microbiol.; 27: 51-61.
• Okamoto K, Misumi Y, Kadowaki T, Yoneda M, Yamamoto K, Ikehara Y. (1995) Structural characterization of Arg gingipain, a novel arginine-specific cysteine proteinase as a major periodontal pathogenic factor from Porphyromonas gingivalis. Arch Biochem Biophys.; 316: 917-25.
• Okamoto K, Kadowaki T, Nakayama K, Yamamoto K. (1996) Cloning and sequencing of the gene encoding a novel lysine-specific cysteine proteinase (Lys-gingipain) in Porphyromonas gingivalis: structural relationship with the arginine-specific cysteine proteinase (Arg-gingipain). J Biochem.; 120: 398-406.
• Okamoto K, Nakayama K, Kadowaki T, Abe N, Ratnayake DB, Yamamoto K. (1998) Involvement of a lysine-specific cysteine proteinase in hemoglobin adsorption and heme accumulation by Porphyromonas gingivalis. J Biol Chem.; 273: 21225-321.
• Olczak T, Dixon DW, Genco CA. (2001) Binding specificity of the Porphyromonas gingivalis heme and hemoglobin receptor HmuR, gingipain K, and gingipain R1 for heme, porphyrins, and metalloporphyrins. J Bacteriol.; 183: 5599-608.
• Paramaesvaran M, Nguyen KA, Caldon E, McDonald JA, Najdi S, Gonzaga G, Langley DB, DeCarlo A, Crossley MJ, Hunter N, Collyer CA. (2003) Porphyrin-mediated cell surface heme capture from hemoglobin by Porphyromonas gingivalis. J Bacteriol.; 185: 2528-37.
• Pavloff N, Potempa J, Pike RN, Prochazka V, Kiefer MC, Travis J, Barr PJ. (1995) Molecular cloning and structural characterization of the Arg-gingipain proteinase of Porphyromonas gingivalis. Biosynthesis as a proteinase-adhesion polyprotein. J Biol Chem.; 270: 1007-10.
• Pavloff N, Pemberton PA, Potempa J, Chen WCA, Pike RN, Prochazka V, Kiefer MC, Travis J, Barr PJ. (1997) Molecular cloning and characterization of Porphyromonas gingivalis Lys-gingipain. A new member of an emerging family of pathogenic bacterial cysteine proteinases. J Biol Chem.; 272: 1595-600.
• Pike RN, McGraw W, Potempa J, Travis J. (1994) Lysine- and arginine-specific proteinases from Porphyromonas gingivalis. Isolation, characterization and evidence for the existence of complexes with hemagglutinins. J Biol Chem.; 269: 406-11.
• Potempa J, Pike R, Travis J. (1997) Titration and mapping of the active site of cysteine proteinases from Porphyromonas gingivalis (gingipains) using peptidyl chloromethanes. Biol Chem.; 378: 223-30.
• Potempa J, Mikolajczyk-Pawlinska J, Brassell D, Nelson D, Thogersen IB, Enghild JJ, Travis J. (1998) Comparative properties of two cysteine proteinases (gingipains R), the products of two related but individual genes of Porphyromonas gingivalis. J Biol Chem.; 273: 21648-57.
• Shi Y, Ratnayake DB, Okamoto K, Abe N, Yamamoto K, Nakayama K. (1999) Genetic analyses of proteolysis, hemoglobin binding, and hemagglutination of Porphyromonas gingivalis. J Biol Chem.; 274: 17955-60.
• Simpson W, Wang CY, Bond V, Potempa J, Mikolajczyk-Pawlinska J, Travis J, Genco CA. (1999) Transposition of the endogenous insertion sequence element IS1126 modulates gingipain expression in Porphyromonas gingivalis. Infect Immun.; 67: 5012-20.
• Simpson W, Olczak T, Genco CA. (2000) Characterization and expression of HmuR, a TonB-dependent hemoglobin receptor of Porphyromonas gingivalis. J Bacteriol.; 182: 5737-48.
• Slakeski N, Dashper SG, Cook P, Poon C, Moore C, Reynolds EC. (2000) A Porphyromonas gingivalis genetic locus encoding a heme transport system. Oral Microbiol Immunol.; 15: 388-92.
• Sroka AE, Sztukowska M, Potempa J, Travis J, Genco CA. (2001) Degradation of host heme proteins by the lysine- and arginine-specific cysteine proteinases (gingipains) of Porphyromonas gingivalis. J Bacteriol.; 183: 5609-16.
• Tokuda M, Chen W, Karunakaran T, Kuramitsu H. (1998) Regulation of protease expression in Porphyromonas gingivalis. Infect Immun.; 66: 5232-7.