ClpL

Drug – bio-affecting and body treating compositions – Enzyme or coenzyme containing – Hydrolases

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Details

424 9463, 435195, 435196, 4352523, 4353201, 435883, 536 232, 536 234, A61K 3846, A61K 3848, C12N 916, C12N 952, C07M 2104

Patent

active

061269379

DESCRIPTION:

BRIEF SUMMARY
FIELD OF THE INVENTION

This invention relates to newly identified polynucleotides and polypeptides and their production and uses, as well as their variants, agonists and antagonists, and their uses. In particular, in these and in other regards, the invention relates to polynucleotides and polypeptides of the Clp ATPases family hereinafter referred to as "clpL".


BACKGROUND OF THE INVENTION

The frequency of Staphylococcus aureus infections has risen dramatically in the past 20 years. This has been attributed to the emergence of multiply antibiotic resistant strains and an increasing population of people with weakened immune systems. It is no longer uncommon to isolate Staphylococcus aureus strains which are resistant to some or all of the standard antibiotics. This has created a demand for both new anti-microbial agents and diagnostic tests for this organism.
The Clp ATPases constitute a recently discovered family that is represented in every organism examined thus far. They appear to serve at least two roles in the cell. They are involved in the regulation of proteolysis by activating the ClpP protease in the ATP-dependent cleavage of denatured proteins. (Squires & Squires, 1992, The Clp Proteins: Proteolysis regulators or molecular chaperones? Journal of Bacteriology 174:1081-5). They are also molecular chaperones which can function independently of ClpP protease, and are capable of repairing proteins damaged during stress conditions (Wawrzynow A., Banecki, B. & Zylicz, M., 1996,. The Clp ATPases define a novel class of molecular chaperones. Molecular Microbiology 21:895-9). Microorganisms such as Staphylococcus aureus are under a variety of environmental stresses in vivo, and it can be expected that members of the Clp ATPase family will play a role in survival and growth of the pathogen during infection. Consistent with this is the discovery of a gene encoding a Clp-like protein which is expressed in vivo. Inhibition of the function of this protein is likely to impair the ability of Staphylococcus aureus to establish and maintain an infection.
Clearly, there is a need for factors, such as the compounds of the invention, that have a present benefit of being useful to screen compounds for antibiotic activity. Such factors are also useful to determine their role in pathogenesis of infection, dysfunction and disease. There is also a need for identification and characterization of such factors and their antagonists and agonists which can play a role in preventing, ameliorating or correcting infections, dysfunctions or diseases.
The polypeptides of the invention have amino acid sequence homology to a known Lactococcus lactis clpL protein.


SUMMARY OF THE INVENTION

It is an object of the invention to provide polypeptides that have been identified as clpL polypeptides of the invention by homology between the amino acid sequence set out in Table 1 [SEQ ID NO: 2] and a known amino acid sequence or sequences of other proteins such as Lactococcus lactis clpL protein.
It is a further object of the invention to provide polynucleotides that encode clpL polypeptides, particularly polynucleotides that encode the polypeptide herein designated clpL.
In a particularly preferred embodiment of the invention, the polynucleotide comprises a region encoding clpL polypeptides comprising the sequence set out in Table 1 [SEQ ID NO: 1], or a variant thereof.
In another particularly preferred embodiment of the invention, there is a clpL protein from Staphylococcus aureus comprising the amino acid sequence of Table 1 [SEQ ID NO:2], or a variant thereof.
In accordance with another aspect of the invention, there is provided an isolated nucleic acid molecule encoding a mature polypeptide expressible by the Staphylococcus aureus WCUH 29 strain contained in the deposited strain.
As a further aspect of the invention, there are provided isolated nucleic acid molecules encoding clpL, particularly Staphylococcus aureus clpL, including mRNAs, cDNAs, genomic DNAs. Further embodiments of the invention include biologically, diagnostically, pr

REFERENCES:
Birkenbihl, et al., "Cloning and characterization of rad21, an essential gene of Schizosaccharomyces pombe involved in DNA double-strand-break repair", Nucleic Acids Research, vol. 20, pp. 6605-6611 (sequence search), 1992.
Huang, et al., "Two genes present on a transposon-like strucutre in Lactococcus lactis are involved in a Clp-family proteolytic activity", Molecular Microbiology, vol. 7, No. 6, pp. 957-965, (1993).
Wawrzynow, et al., "The Clp ATPases define a novel class of molecular chaperones", Molecular Microbiology, vol. 21, pp. 895-899, May, 1996.
Lazazzera, et al., "A regulatory switch involving a Clp ATPase", BioEssays, vol. 19, No. 6, pp. 455-458, Jun., 1997.
Seol, et al., "Distinctive Roles of the Two ATP-binding Sites in Clpa, the ATPase Component of Protease It in Escherichia coli", The Journal of Biological Chemistry, vol. 270, pp. 8087-8092, Apr. 7, 1995.

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