The purification and characterization of an 88-kDa Porphyromonas endodontalis 35406 protease
A Porphyromonas endodontalis ATCC 35406 protease was purified from Triton X-114 cell extracts by preparative SDS-PAGE followed by electroelution. The purified enzyme exhibits a molecular size of 88 kDa and was dissociated into two polypeptides of 43 and 41 kDa upon heating in the presence of sodium dodecyl sulfate with or without a reducing agent. The protease (pH optimum 7.5-8.0) degraded the extracellular matrix proteins fibrinogen and fibronectin. Collagen IV was also degraded at 37 degrees C but not at 28 degrees C. The protease also cleaved the bioactive peptide angiotensin at amino acid residue phenylalanine-8 and tyrosine-4 but failed to hydrolyze bradykinin, vasopressin and synthetic chromogenic substrates with phenylalanine or tyrosine at the P1 position. In addition, two peptidases were detected in P. endodontalis cells: a proline aminopeptidase that remained associated with the cell pellet after detergent extraction and peptidase/s that partitioned into the Triton X-114 phase after phase separation and degraded the bioactive peptides bradykinin and vasopressin. These P. endodontalis peptidases and proteases may play an important role in both the nutrition and pathogenicity of these assacharolytic microorganisms. The inactivation of bioactive peptides and degradation of extracellular matrix proteins by bacterial enzymes may contribute to the damage of host tissues accompanied with endodontic infections.
| Medienart: |
Artikel |
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| Erscheinungsjahr: |
2001 |
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| Erschienen: |
2001 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:16 |
|---|---|
| Enthalten in: |
Oral microbiology and immunology - 16(2001), 6 vom: 15. Dez., Seite 326-31 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Rosen, G [VerfasserIn] |
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| Anmerkungen: |
Date Completed 08.02.2002 Date Revised 25.10.2019 published: Print Citation Status MEDLINE |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM116285729 |
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| LEADER | 01000naa a22002652 4500 | ||
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| 003 | DE-627 | ||
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| 007 | tu | ||
| 008 | 231222s2001 xx ||||| 00| ||eng c | ||
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| 035 | |a (DE-627)NLM116285729 | ||
| 035 | |a (NLM)11737654 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Rosen, G |e verfasserin |4 aut | |
| 245 | 1 | 4 | |a The purification and characterization of an 88-kDa Porphyromonas endodontalis 35406 protease |
| 264 | 1 | |c 2001 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ohne Hilfsmittel zu benutzen |b n |2 rdamedia | ||
| 338 | |a Band |b nc |2 rdacarrier | ||
| 500 | |a Date Completed 08.02.2002 | ||
| 500 | |a Date Revised 25.10.2019 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a A Porphyromonas endodontalis ATCC 35406 protease was purified from Triton X-114 cell extracts by preparative SDS-PAGE followed by electroelution. The purified enzyme exhibits a molecular size of 88 kDa and was dissociated into two polypeptides of 43 and 41 kDa upon heating in the presence of sodium dodecyl sulfate with or without a reducing agent. The protease (pH optimum 7.5-8.0) degraded the extracellular matrix proteins fibrinogen and fibronectin. Collagen IV was also degraded at 37 degrees C but not at 28 degrees C. The protease also cleaved the bioactive peptide angiotensin at amino acid residue phenylalanine-8 and tyrosine-4 but failed to hydrolyze bradykinin, vasopressin and synthetic chromogenic substrates with phenylalanine or tyrosine at the P1 position. In addition, two peptidases were detected in P. endodontalis cells: a proline aminopeptidase that remained associated with the cell pellet after detergent extraction and peptidase/s that partitioned into the Triton X-114 phase after phase separation and degraded the bioactive peptides bradykinin and vasopressin. These P. endodontalis peptidases and proteases may play an important role in both the nutrition and pathogenicity of these assacharolytic microorganisms. The inactivation of bioactive peptides and degradation of extracellular matrix proteins by bacterial enzymes may contribute to the damage of host tissues accompanied with endodontic infections | ||
| 650 | 4 | |a Journal Article | |
| 650 | 7 | |a Angiotensins |2 NLM | |
| 650 | 7 | |a Chromogenic Compounds |2 NLM | |
| 650 | 7 | |a Collagen Type IV |2 NLM | |
| 650 | 7 | |a Fibronectins |2 NLM | |
| 650 | 7 | |a Oxidants |2 NLM | |
| 650 | 7 | |a Surface-Active Agents |2 NLM | |
| 650 | 7 | |a Vasopressins |2 NLM | |
| 650 | 7 | |a 11000-17-2 |2 NLM | |
| 650 | 7 | |a Sodium Dodecyl Sulfate |2 NLM | |
| 650 | 7 | |a 368GB5141J |2 NLM | |
| 650 | 7 | |a Tyrosine |2 NLM | |
| 650 | 7 | |a 42HK56048U |2 NLM | |
| 650 | 7 | |a Phenylalanine |2 NLM | |
| 650 | 7 | |a 47E5O17Y3R |2 NLM | |
| 650 | 7 | |a Fibrinogen |2 NLM | |
| 650 | 7 | |a 9001-32-5 |2 NLM | |
| 650 | 7 | |a Endopeptidases |2 NLM | |
| 650 | 7 | |a EC 3.4.- |2 NLM | |
| 650 | 7 | |a Aminopeptidases |2 NLM | |
| 650 | 7 | |a EC 3.4.11.- |2 NLM | |
| 650 | 7 | |a Cystinyl Aminopeptidase |2 NLM | |
| 650 | 7 | |a EC 3.4.11.3 |2 NLM | |
| 650 | 7 | |a prolyl aminopeptidase |2 NLM | |
| 650 | 7 | |a EC 3.4.11.5 |2 NLM | |
| 650 | 7 | |a Lysine Carboxypeptidase |2 NLM | |
| 650 | 7 | |a EC 3.4.17.3 |2 NLM | |
| 650 | 7 | |a Bradykinin |2 NLM | |
| 650 | 7 | |a S8TIM42R2W |2 NLM | |
| 700 | 1 | |a Shoshani, M |e verfasserin |4 aut | |
| 700 | 1 | |a Naor, R |e verfasserin |4 aut | |
| 700 | 1 | |a Sela, M N |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Oral microbiology and immunology |d 1987 |g 16(2001), 6 vom: 15. Dez., Seite 326-31 |w (DE-627)NLM012649619 |x 0902-0055 |7 nnns |
| 773 | 1 | 8 | |g volume:16 |g year:2001 |g number:6 |g day:15 |g month:12 |g pages:326-31 |
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| 951 | |a AR | ||
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