DOI: 10.1046/j.1432-1327.1999.00065.x. Domain structure, GTP‐hydrolyzing activity and 7S RNA binding of <i>Acidianus ambivalens</i> Ffh‐homologous protein suggest an SRP‐like complex in archaea

Domain structure, GTP‐hydrolyzing activity and 7S RNA binding of Acidianus ambivalens Ffh‐homologous protein suggest an SRP‐like complex in archaea

10.1046/j.1432-1327.1999.00065.x

Crossref journal-article

Wiley

European Journal of Biochemistry (311)

Abstract

In this study we provide, for the first time, experimental evidence that a protein homologous to bacterial Ffh is part of an SRP‐like ribonucleoprotein complex in hyperthermophilic archaea. The gene encoding the Ffh homologue in the hyperthermophilic archaeote Acidianus ambivalens has been cloned and sequenced. Recombinant Ffh protein was expressed in E. coli and subjected to biochemical and functional studies. A. ambivalens Ffh encodes a 50.4‐kDa protein that is structured by three distinct regions: the N‐terminal hydrophilic N‐region (N), the GTP/GDP‐binding domain (G) and a C‐terminal located C‐domain (C). The A. ambivalens Ffh sequence shares 44–46% sequence similarity with Ffh of methanogenic archaea, 34–36% similarity with eukaryal SRP54 and 30–34% similarity with bacterial Ffh. A polyclonal antiserum raised against the first two domains of A. ambivalens Ffh reacts specifically with a single protein (apparent molecular mass: 46 kDa, termed p46) present in cytosolic and in plasmamembrane cell fractions of A. ambivalens. Recombinant Ffh has a melting point of tm = 89 °C. Its intrinsic GTPase activity obviously depends on neutral pH and low ionic strength with a preference for chloride and acetate salts. Highest rates of GTP hydrolysis have been achieved at 81 °C in presence of 0.1–1 mm Mg2+. GTP hydrolysis is significantly inhibited by high glycerol concentrations, and the GTP hydrolysis rate also markedly decreases by addition of detergents. The Km for GTP is 13.7 µm at 70 °C and GTP hydrolysis is strongly inhibited by GDP (Ki = 8 µm). A. ambivalens Ffh, which includes an RNA‐binding motif in the C‐terminal domain, is shown to bind specifically to 7S RNA of the related crenarchaeote Sulfolobus solfataricus. Comparative sequence analysis reveals the presence of typical signal sequences in plasma membrane as well as extracellular proteins of hyperthermophilic crenarchaea which strongly supposes recognition events by an Ffh containing SRP‐like particle in these organisms.

Bibliography

Moll, R., Schmidtke, S., & SchÃ¤fer, G. (1999). Domain structure, GTPâhydrolyzing activity and 7S RNA binding of Acidianus ambivalens Ffhâhomologous protein suggest an SRPâlike complex in archaea. European Journal of Biochemistry, 259(1â2), 441â448. Portico.

Authors 3

Ralf Moll (first)
Silke Schmidtke (additional)
Günter Schäfer (additional)

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Dates

Type	When
Created	22 years, 5 months ago (March 11, 2003, 12:12 p.m.)
Deposited	2 years ago (Aug. 31, 2023, 8:04 a.m.)
Indexed	2 months ago (July 3, 2025, 12:03 p.m.)
Issued	26 years, 8 months ago (Jan. 1, 1999)
Published	26 years, 8 months ago (Jan. 1, 1999)
Published Online	23 years, 8 months ago (Dec. 25, 2001)
Published Print	26 years, 8 months ago (Jan. 1, 1999)

Funders 0

None

BibTeX

@article{Moll_1999, title={Domain structure, GTP‐hydrolyzing activity and 7S RNA binding of Acidianus ambivalens Ffh‐homologous protein suggest an SRP‐like complex in archaea}, volume={259}, ISSN={1432-1033}, url={http://dx.doi.org/10.1046/j.1432-1327.1999.00065.x}, DOI={10.1046/j.1432-1327.1999.00065.x}, number={1–2}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={Moll, Ralf and Schmidtke, Silke and Schäfer, Günter}, year={1999}, month=jan, pages={441–448} }

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