53010, a human carboxylesterase family member and uses thereof

Details 53010, a human carboxylesterase family member and uses thereof 53010, a human carboxylesterase family member and uses thereof

2001-12-18

2003-12-16

Prouty, Rebecca E. (Department: 1652)

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Hydrolase

C435S252300, C435S252330, C435S320100, C435S325000, C536S023200

Reexamination Certificate

active

06664091

ABSTRACT:

BACKGROUND OF THE INVENTION
Higher eukaryotes have many distinct esterases. Among the different types of esterases are those that act on carboxylic esters, also known as carboxylesterases. Carboxylesterases have been classified into three categories (A, B and C) on the basis of differential patters of inhibition by organophosphates (Myers et al. (1988)
Mol. Biol. Evol.
5(2):113-119). Sequence analysis of a number of type-B carboxylesterase demonstrates their evolutionary interrelatedness. Members of the type B carboxylesterase family include acetylcholincarboxylesterases, mammalian cholincarboxylesterases, thyroglobulin, neuroligins, and mammalian bile salt activated lipases.
The type B family of carboxylesterase also includes vitamin K-dependent carboxylases. Vitamin K-dependent gamma-glutamyl carboxylases catalyze the posttranslational conversion of glutamic acid to gamma-carboxyglutamic acid, an amino acid critical to the function of the vitamin K-dependent blood coagulation proteins (Begley et al. (2000)
J. Biol. Chem.
275:36245-36249). Incomplete gamma carboxylation of blood clotting factors is associated with poor coagulation.
For the gamma carboxylation event to occur, both vitamin K and the presence of a gamma carboxylation recognition site on the substrate are required. Gamma carboxyglutamic acid confers calcium binding ability upon the modified protein. For blood clotting factors, calcium binding results in a conformational change that exposes hydrophobic residues for interactions with membranes.
Although gamma carboxylation was a biochemical event first characterized in the mammalian blood clotting cascade, it has been found to have a more generalized applicability. For example, vitamin K-dependent gamma carboxylation of glutamate residues has also been detected for a variety of other proteins including bone proteins, PRGP1, PRGP2, and neuropeptides (Walker et al. (2000)
J. Biol. Chem., December
7 epub ahead of print).
SUMMARY OF THE INVENTION
The present invention is based, in part, on the discovery of a novel carboxylesterase family member, referred to herein as “53010.” The nucleotide sequence of a cDNA encoding 53010 is shown in SEQ ID NO:1, and the amino acid sequence of a 53010 polypeptide is shown in SEQ ID NO:2. In addition, the nucleotide sequences of the coding region are depicted in SEQ ID NO:3.
Accordingly, in one aspect, the invention features a nucleic acid molecule that encodes a 53010 protein or polypeptide, e.g., a biologically active portion of the 53010 protein. In a preferred embodiment the isolated nucleic acid molecule encodes a polypeptide having the amino acid sequence of SEQ ID NO:2. In other embodiments, the invention provides isolated 53010 nucleic acid molecules having the nucleotide sequence shown in SEQ ID NO:1, SEQ ID NO:3, a full complement of SEQ ID NO:1 or SEQ ID NO:3. In still other embodiments, the invention provides nucleic acid molecules that are substantially identical (e.g., naturally occurring allelic variants) to the nucleotide sequence shown in SEQ ID NO:1, SEQ ID NO:3. In other embodiments, the invention provides a nucleic acid molecule which hybridizes under a stringency condition described herein to a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:3, wherein the nucleic acid encodes a full length 53010 protein or an active fragment thereof.
In a related aspect, the invention further provides nucleic acid constructs that include a 53010 nucleic acid molecule described herein. In certain embodiments, the nucleic acid molecules of the invention are operatively linked to native or heterologous regulatory sequences. Also included, are vectors and host cells containing the 53010 nucleic acid molecules of the invention e.g., vectors and host cells suitable for producing 53010 nucleic acid molecules and polypeptides.
In another related aspect, the invention provides nucleic acid fragments suitable as primers or hybridization probes for the detection of 53010-encoding nucleic acids.
In still another related aspect, isolated nucleic acid molecules that are antisense to a 53010 encoding nucleic acid molecule are provided.
In another aspect, the invention features, 53010 polypeptides, and biologically active or antigenic fragments thereof that are useful, e.g., as reagents or targets in assays applicable to treatment and diagnosis of 53010-mediated or -related disorders. In another embodiment, the invention provides 53010 polypeptides having a 53010 activity. Preferred polypeptides are 53010 proteins including at least one carboxylesterase domain, and, preferably, having a 53010 activity, e.g., a 53010 activity as described herein.
In other embodiments, the invention provides 53010 polypeptides, e.g., a 53010 polypeptide having the amino acid sequence shown in SEQ ID NO:2 or; an amino acid sequence that is substantially identical to the amino acid sequence shown in SEQ ID NO:2; or an amino acid sequence encoded by a nucleic acid molecule having a nucleotide sequence which hybridizes under a stringency condition described herein to a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:3, wherein the nucleic acid encodes a full length 53010 protein or an active fragment thereof.
In a related aspect, the invention further provides nucleic acid constructs which include a 53010 nucleic acid molecule described herein.
In a related aspect, the invention provides 53010 polypeptides or fragments operatively linked to non-53010 polypeptides to form fusion proteins.
In another aspect, the invention features antibodies and antigen-binding fragments thereof, that react with, or more preferably specifically bind 53010 polypeptides or fragments thereof, e.g., a carboxylesterase domain.
In another aspect, the invention provides methods of screening for agents, e.g., compounds, that modulate the expression or activity of the 53010 polypeptides or nucleic acids, e.g., compounds that modulate the normal pain response, aberrant or altered pain response, or neurological response.
In a preferred embodiment, the effect of an agent, e.g., a compound, on the pain response is evaluated by an analgesic test, e.g., the hot plate test, tail flick test, writhing test, paw pressure test, all electric stimulation test, tail withdrawal test, or formalin test.
In a preferred embodiment, the agent, e.g., compound, inhibits 53010 activity.
In still another aspect, the invention provides a process for modulating 53010 polypeptide or nucleic acid expression or activity, e.g. using the screened compounds. In certain embodiments, the methods involve treatment of conditions related to aberrant, e.g., decreased or increased expression of the 53010 polypeptides or nucleic acids, such as conditions involving pain response, aberrant or altered pain response, or pain related disorders.
In still another aspect, the invention features a method of modulating (e.g., enhancing or inhibiting) an activity of a cell (e.g., a neural cell), or a response (e.g., a pain response) in a subject. The method includes contacting the cell with, or administered to the subject, an agent, e.g., a compound, that modulates the activity or expression of a 53010 polypeptide or nucleic acid, in an amount effective to modulate the activity or the response.
In a preferred embodiment, the agent modulates (e.g., increases or decreases) carboxylesterase activity.
In a preferred embodiment, the agent modulates (e.g., increases or decreases) expression of the 53010 nucleic acid by, e.g., modulating transcription, mRNA stability, etc.
In a preferred embodiment, the cell, e.g., the 53010-expressing cell, is a central or peripheral nervous system cell, e.g., a cell in an area involved in pain control, e.g., a cell in the brain or spinal cord.
In a preferred embodiment, the agent, e.g., the compound, and the 53010-polypeptide or nucleic acid are contacted in vitro or ex vivo. In a preferred embodiment, the contacting step is effected in vivo in a subject, e.g., as part of a therapeutic or prophylactic protocol. The contacting or administerin

Affiliated with

Also associated with

Rating

53010, a human carboxylesterase family member and uses thereof does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with 53010, a human carboxylesterase family member and uses thereof, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and 53010, a human carboxylesterase family member and uses thereof will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3169636