Glycogen phosphorylase from streptococcus pneumoniae

Details Glycogen phosphorylase from streptococcus pneumoniae Glycogen phosphorylase from streptococcus pneumoniae

1998-11-04

2001-05-22

Duffy, Patricia A. (Department: 1645)

Drug, bio-affecting and body treating compositions

Antigen, epitope, or other immunospecific immunoeffector

Fusion protein or fusion polypeptide

C424S190100, C424S244100, C435S195000, C435S200000

Reexamination Certificate

active

06235285

ABSTRACT:

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 novel polynucleotides and polypeptides of the glycogen phosphorylase family, hereinafter referred to as “glycogen phosphorylase”.
BACKGROUND OF THE INVENTION
The Streptococci make up a medically important genera of microbes known to cause several types of disease in humans, including, for example, otitis media, conjunctivitis, pneumonia, bacteremia, meningitis, sinusitis, pleural empyema and endocarditis, and most particularly meningitis, such as for example infection of cerebrospinal fluid. Since its isolation more than 100 years ago,
Streptococcus pneumoniae
has been one of the more intensively studied microbes. For example, much of our early understanding that DNA is, in fact, the genetic material was predicated on the work of Griffith and of Avery, Macleod and McCarty using this microbe. Despite the vast amount of research with
S. pneumoniae,
many questions concerning the virulence of this microbe remain. It is particularly preferred to employ Streptococcal genes and gene products as targets for the development of antibiotics.
The frequency of
Streptococcus pneumoniae
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
Streptococcus pneumoniae
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.
Glycogen phosphorylase is a catabolic gene involved in the breakdown of glycogen, an energy storage carbohydrate in many organisms. It has been detected in a wide range of bacteria (Romeo T, Kumar A, Preiss J Gene Oct. 30, 1988; 70(2):363-376, Kiel J A, Boels J M, Beldman G, Venema G Mol Microbiol Jan. 11, 1994(l):203-218 and Preiss J, Romeo T Adv Microb Physiol 1989, 30:183-238) The breakdown of glycogen may be particularly important in adverse situations for bacterial growth such as infection in mammals.
Clearly, there is a need for factors, such as the novel 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 maltodextrin phosphorylase (phsm_ecoli) protein.
SUMMARY OF THE INVENTION
It is an object of the invention to provide polypeptides that have been identified as novel glycogen phosphorylase polypeptides 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 MALTODEXTRIN PHOSPHORYLASE (PHSM_ECOLI) protein.
It is a further object of the invention to provide polynucleotides that encode glycogen phosphorylase polypeptides, particularly polynucleotides that encode the polypeptide herein designated glycogen phosphorylase.
In a particularly preferred embodiment of the invention the polynucleotide comprises a region encoding glycogen phosphorylase polypeptides comprising the sequence set out in Table 1 [SEQ ID NO:1] which includes a full length gene, or a variant thereof.
In another particularly preferred embodiment of the invention there is a novel glycogen phosphorylase protein from
Streptococcus pneumoniae
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
Streptococcus pneumoniae
0100993 strain contained in the deposited strain.
A further aspect of the invention there are provided isolated nucleic acid molecules encoding glycogen phosphorylase, particularly
Streptococcus pneumoniae
glycogen phosphorylase, including mRNAs, cDNAs, genomic DNAs. Further embodiments of the invention include biologically, diagnostically, prophylactically, clinically or therapeutically useful variants thereof, and compositions comprising the same.
In accordance with another aspect of the invention, there is provided the use of a polynucleotide of the invention for therapeutic or prophylactic purposes, in particular genetic immunization. Among the particularly preferred embodiments of the invention are naturally occurring allelic variants of glycogen phosphorylase and polypeptides encoded thereby.
Another aspect of the invention there are provided novel polypeptides of
Streptococcus pneumoniae
referred to herein as glycogen phosphorylase as well as biologically, diagnostically, prophylactically, clinically or therapeutically useful variants thereof, and compositions comprising the same.
Among the particularly preferred embodiments of the invention are variants of glycogen phosphorylase polypeptide encoded by naturally occurring alleles of the glycogen, phosphorylase genie.
In a preferred embodiment of the invention there are provided methods for producing the aforementioned glycogen phosphorylase polypeptides.
In accordance with yet another aspect of the invention, there are provided inhibitors to such polypeptides, useful as antibacterial agents, including, for example, antibodies.
In accordance with certain preferred embodiments of the invention, there are provided products, compositions and methods for assessing glycogen phosphorylase expression, treating disease, for example, otitis media conjunctivitis, pneumonia, bacteremia, meningitis, sinusitis, pleural empyema and endocarditis, and most particularly meningitis, such as for example infection of cerebrospinal fluid, assaying genetic variation, and administering a glycogen phosphorylase polypeptide or polynucleotide to an organism to raise an immunological response against a bacteria, especially a
Streptococcus pneumoniae
bacteria.
In accordance with certain preferred embodiments of this and other aspects of the invention there are provided polynucleotides that hybridize to glycogen phosphorylase polynucleotide sequences, particularly under stringent conditions.
In certain preferred embodiments of the invention there are provided antibodies against glycogen phosphorylase polypeptides.
In other embodiments of the invention there are provided methods for identifying compounds which bind to or otherwise interact with and inhibit or activate an activity of a polypeptide or polynucleotide of the invention comprising: contacting a polypeptide or polynucleotide of the invention with a compound to be screened under conditions to permit binding to or other interaction between the compound and the polypeptide or polynucleotide to assess the binding to or other interaction with the compound, such binding or interaction being associated with a second component capable of providing a detectable signal in response to the binding or interaction of the polypeptide or polynucleotide with the compound; and determining whether the compound binds to or otherwise interacts with and activates or inhibits an activity of the polypeptide or polynucleotide by detecting the presence or absence of a signal generated from the binding or interaction of the compound with the polypeptide or polynucleotide.
In accordance with yet another aspect of the invention, there are provided glycogen phosphorylase agonists and antagonists, preferably bacteriostatic or bactericidal agonists and antagonists.
In a further aspect of the invention there are provided compositions comprising a glycogen phosphorylase polynucleotide or a glycogen ph

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