Abstract
CanA from Pyrodictium abyssi forms a heat-resistant organic hollow-fiber network together with CanB and CanC. An N-terminally truncated construct of CanA (K1-CanA) gave NMR spectra of good quality that could be assigned by three-dimensional NMR methods on 15N and 13C–15N enriched protein. We assigned the chemical shifts of 96% of all backbone 1HN atoms, 98% of all backbone 15N atoms, 100% of all 13Cα atoms, 100% of all 1Hα atoms, 90% of all 13C′ atoms, and 100% of the 13Cβ atoms. Two short helices and 10 β-strands are estimated from an analysis of the chemical shifts leading to a secondary structure content of K1-CanA of 6% helices, 44% β-pleated sheets, and 50% coils.
Highlights
The members of the archaeal family Pyrodictiaceae have typical growth temperatures between 75 and 110 °C (348 to 383 K) (Stetter 1982; Stetter et al 1983; Pley et al 1991)
P. abyssi was isolated from a black smoker, a hydrothermal deep-sea vent 3600 m below sea level (Deininger 1994)
This truncated version of CanA has been selected for assignments after limited proteolysis experiments
Summary
The members of the archaeal family Pyrodictiaceae have typical growth temperatures between 75 and 110 °C (348 to 383 K) (Stetter 1982; Stetter et al 1983; Pley et al 1991). Three different species of Pyrodictium are known, P. occultum, P. brockii and P. abyssi. P. abyssi was isolated from a black smoker, a hydrothermal deep-sea vent 3600 m below sea level (Deininger 1994). Form stable tubules with the same characteristics as native cannulae. The heat resistance of the hollow-fibers promises interesting biotechnological applications. Carbon nanotubes are a central research topic. CanA nanotubes could take a similar role in nanobiophysics, since they are heat stable, self-organizing, and can be modified by site-directed mutagenesis. A prerequisite for the structure determination of CanA is the complete sequential assignment of uniformly 13C–15N enriched protein that is presented here
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