A hyperthermostable protease of the subtilisin family bound to the surface layer of the Archaeon Staphylothermus marinus

Mayr, Jutta; Lupas, Andrei N.; Kellermann, Josef; Eckerskorn, Christoph; Baumeister, Wolfgang; Peters, Jürgen

doi:10.1016/s0960-9822(09)00455-2

Cited by 65 publications

(41 citation statements)

References 42 publications

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“…The S-layer protein of H. halobium shows a hydrophobic stretch of 21 amino acids on its C-terminal part which integrates into the plasma membrane (69). A similar observation was reported for the S-layer protein of Staphylothermus marinus, which carries an extremely stable protease on its 70-nm-long stalk (80,99).…”

Section: Attachment Of S-layer Proteins To the Underlying Cell Envelosupporting

confidence: 76%

“…Most S-layers are 5 to 25 nm thick, and they reveal a rather smooth outer surface and a more corrugated inner surface. Among S-layer lattices of archaea, pillar-like extensions on the inner surface can be observed (80,99,100,103). Since S-layers are monomolecular assemblies of identical subunits, they exhibit pores identical in size and morphology.…”

Section: Occurrence Structure and Assemblymentioning

confidence: 99%

“…The largest pores in the 5-to 8-nm range were found in S-layer lattices of those archaea lacking a rigid cell wall layer in which these crystalline arrays fulfill a shape-determining or shape-maintaining function (3,4,50,80,99). As a general feature, the S-layer subunits of archaea lacking a rigid cell wall layer possess long, hydrophobic protrusions which integrate into the plasma membrane.…”

Section: Functional Aspectsmentioning

confidence: 99%

“…Furthermore, two copies of a hyperthermostable protease showing significant similarity to subtilisin are attached near the middle of each stalk. This enzyme is resistant to heat inactivation to 95°C in the free form and to 125°C in the stalk-bound form (80,99).…”

Section: Functional Aspectsmentioning

confidence: 99%

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S-Layer Proteins

2000

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Section: Attachment Of S-layer Proteins To the Underlying Cell Envelosupporting

confidence: 76%

Section: Occurrence Structure and Assemblymentioning

confidence: 99%

Section: Functional Aspectsmentioning

confidence: 99%

Section: Functional Aspectsmentioning

confidence: 99%

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S-Layer Proteins

2000

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“…4). Thus, like other subtilases (25), T. kodakaraensis subtilisin shows a broad substrate specificity with a slight preference to large hydrophobic amino acid residues at the P1 position.…”

Section: Resultsmentioning

confidence: 86%

Active Subtilisin-Like Protease from a Hyperthermophilic Archaeon in a Form with a Putative Prosequence

Kannan¹,

Koga²,

Inoue³

et al. 2001

Appl Environ Microbiol

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The gene encoding subtilisin-like protease T. kodakaraensis subtilisin was cloned from a hyperthermophilic archaeon Thermococcus kodakaraensis KOD1. T. kodakaraensis subtilisin is a member of the subtilisin family and composed of 422 amino acid residues with a molecular weight of 43,783. It consists of a putative presequence, prosequence, and catalytic domain. Like bacterial subtilisins, T. kodakaraensis subtilisin was overproduced in Escherichia coli in a form with a putative prosequence in inclusion bodies, solubilized in the presence of 8 M urea, and refolded and converted to an active molecule. However, unlike bacterial subtilisins, in which the prosequence was removed from the catalytic domain by autoprocessing upon refolding, T. kodakaraensis subtilisin was refolded in a form with a putative prosequence. This refolded protein of recombinant T. kodakaraensis subtilisin which is composed of 398 amino acid residues (Gly ؊82 to Gly 316 ), was purified to give a single band on a sodium dodecyl sulfate (SDS)-polyacrylamide gel and characterized for biochemical and enzymatic properties. The good agreement of the molecular weights estimated by SDSpolyacrylamide gel electrophoresis (44,000) and gel filtration (40,000) suggests that T. kodakaraensis subtilisin exists in a monomeric form. T. kodakaraensis subtilisin hydrolyzed the synthetic substrate N-succinyl-Ala-AlaPro-Phe-p-nitroanilide only in the presence of the Ca 2؉ ion with an optimal pH and temperature of pH 9.5 and 80°C. Like bacterial subtilisins, it showed a broad substrate specificity, with a preference for aromatic or large nonpolar P1 substrate residues. However, it was much more stable than bacterial subtilisins against heat inactivation and lost activity with half-lives of >60 min at 80°C, 20 min at 90°C, and 7 min at 100°C.

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Kristalline Zelloberflächen-Schichten prokaryotischer Organismen (S-Schichten): von der supramolekularen Zellstruktur zur Biomimetik und Nanotechnologie

et al. 1999

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Ein erstaunlich breites Anwendungspotential in der Biotechnologie, Biomimetik und Nanotechnologie ergaben Untersuchungen von Struktur, Chemie, Biosynthese, Genetik, Morphogenese und Funktion supramolekularer S‐Schicht‐Gitter („S”︁ für surface). Bei diesen handelt es sich um monomolekulare, kristalline Anordnungen aus Protein‐ oder Glycoprotein‐Untereinheiten; sie zählen zu den häufigsten Oberflächenstrukturen von prokaryotischen Zellen (siehe schematische Darstellung der Zellhülle eines Archaebakteriums).

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A hyperthermostable protease of the subtilisin family bound to the surface layer of the Archaeon Staphylothermus marinus

Cited by 65 publications

References 42 publications

S-Layer Proteins

S-Layer Proteins

Active Subtilisin-Like Protease from a Hyperthermophilic Archaeon in a Form with a Putative Prosequence

Kristalline Zelloberflächen-Schichten prokaryotischer Organismen (S-Schichten): von der supramolekularen Zellstruktur zur Biomimetik und Nanotechnologie

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