Difference between revisions of "Part:BBa K2710000"

Revision as of 03:55, 16 October 2018

His-Alpha Prefoldin

alpha-Prefoldin with His-Tag

Usage and Biology

Prefoldin is a hetero-hexameric chaperone protein consisting of two α and four β subunits1, which was isolated from the thermophilic bacteria Methanobacterium thermoautotrophicum². As a result, prefoldin is stable at high temperatures, with a Tm ≥ 70°C². This 85kDa molecular chaperone protein is self-assembling, and consists of six α-helix coiled-coils (with each subunit as a coiled coil) protruding from a double β-barrel³. Each α subunit forms two β hairpins, and the four β hairpins from the two α subunits forms the two 8-stranded up and down β barrels². Additionally, each β subunit forms one β hairpin. The subunits are anchored to the barrel via their proximal ends, and are fully solvated². The α subunits are 85 Angstrom in height, and the β subunits are 70 Angstrom in height, with an overall distance of 50 Angstrom across the double β-barrel². Sigert’s research also suggests that the coiled coils are flexible, although the overall structure is highly conserved². Prefoldin is a molecular chaperone which, in archaea, carries out ‘de novo protein folding’, and in eukaryotes assists in the biogenesis of cytoskeletal proteins, thanks to its coiled coil ‘tentacles’¹. We, however, are using it as the backbone to our modular, thermostable enzyme scaffold.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 <h3>Usage and Biology</h3>
-<p>Prefoldin is a hetero-hexameric protein consisting of two α and four β subunits1, which was isolated from the thermophilic bacteria <i>Methanobacterium thermoautotrophicum</i><sup>2</sup>. As a result, prefoldin is stable at high temperatures, with a Tm ≥ 70°C<sup>2</sup>. This 85kDa molecular chaperone protein is self-assembling, and consists of six α-helix coiled-coils (with each subunit as a coiled coil) protruding from a double β-barrel<sup>3</sup>. Each α subunit forms two β hairpins, and the four β hairpins from the two α subunits forms the two 8-stranded up and down β barrels<sup>2</sup>. Additionally, each β subunit forms one β hairpin. The subunits are anchored to the barrel via their proximal ends, and are fully solvated<sup>2</sup>.
+<p>Prefoldin is a hetero-hexameric chaperone protein consisting of two α and four β subunits1, which was isolated from the thermophilic bacteria <i>Methanobacterium thermoautotrophicum</i><sup>2</sup>. As a result, prefoldin is stable at high temperatures, with a Tm ≥ 70°C<sup>2</sup>. This 85kDa molecular chaperone protein is self-assembling, and consists of six α-helix coiled-coils (with each subunit as a coiled coil) protruding from a double β-barrel<sup>3</sup>. Each α subunit forms two β hairpins, and the four β hairpins from the two α subunits forms the two 8-stranded up and down β barrels<sup>2</sup>. Additionally, each β subunit forms one β hairpin. The subunits are anchored to the barrel via their proximal ends, and are fully solvated<sup>2</sup>.
 The α subunits are 85 Angstrom in height, and the β subunits are 70 Angstrom in height, with an overall distance of 50 Angstrom across the double β-barrel<sup>2</sup>. Sigert’s research also suggests that the coiled coils are flexible, although the overall structure is highly conserved<sup>2</sup>. Prefoldin is a molecular chaperone which, in archaea, carries out ‘de novo protein folding’, and in eukaryotes assists in the biogenesis of cytoskeletal proteins, thanks to its coiled coil ‘tentacles’<sup>1</sup>. We, however, are using it as the backbone to our modular, thermostable enzyme scaffold.</p>