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Antibodies

a technology of nucleic acids and antibodies, applied in the field of nucleic acids coding, can solve the problems of difficult acquisition of large amounts of pure protein, insufficient information, complicated characterization of a particular type of calcium channel by analysis of whole cells, etc., and achieve the effect of modulating the proliferation and growth of these cells and reducing extracellular calcium influx

Inactive Publication Date: 2007-02-08
BETH ISRAEL DEACONESS MEDICAL CENT INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

Enables the modulation of calcium transport and provides therapeutic agents for conditions related to aberrant calcium transport, as well as tools for delivering agents to specific cells, such as hematopoietic cells, and potential targets for designing inhibitors to regulate lymphocyte proliferation.

Problems solved by technology

Characterization of a particular type of calcium channel by analysis of whole cells is complicated by the presence of mixed populations of different types of calcium channels in the majority of cells.
Although single-channel recording methods can be used to examine individual calcium channels, such analysis does not reveal information related to the molecular structure or biochemical composition of the channel.
However, acquiring large amounts of pure protein is difficult in view of the complex nature of these multisubunit proteins, the varying concentrations of calcium channel proteins in tissue sources, the presence of mixed populations of calcium channel proteins in tissues, and the modifications of the native protein that can occur during the isolation procedure.

Method used

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  • Antibodies

Examples

Experimental program
Comparison scheme
Effect test

example 1

Transient Expression of SOC / CRAC

[0191] In the above-identified cell lines and using both of the foregoing expression techniques, SOC / CRAC expression enhances thapsigargin-dependent influx. In addition, SOC / CRAC expression also enhances the amount of intracellular calcium stores. That this effect is likely due to SOC / CRAC acting as a plasma membrane calcium channel can be confirmed by producing an in-frame carboxy-terminal translational fusion with green fluorescent protein followed by confocal microscopy, revealing that SOC / CRAC is expressed predominantly as a plasma membrane calcium channel.

example 2

Patch Clamp Analysis

[0192] The biophysical characteristics of SOC / CRAC enhanced currents when expressed in Xenopus oocytes are determined. SOC / CRAC cRNA injection is able to enhance thapsigargin-dependent whole cell currents. In addition, SOC / CRAC does not alter the reversal potential of these currents and the determination of the Pca / PNa ratio shows that SOC / CRAC channels are highly calcium selective.

example 3

Pharmacologic Behavior of SOC / CRAC

[0193] The pharmacologic behavior of SOC / CRAC is evaluated as described above. SOC / CRAC-enhanced influx is inhibited by sphingosine in a manner that is substantially the same as that of endogenous thapsigargin-dependent calcium influx.

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Abstract

Nucleic acids encoding SOC / CRAC calcium channel polypeptides, including fragments and biologically functional variants thereof and encoded polypeptides are provided. The nucleic acids and polypeptides disclosed herein are useful as therapeutic and diagnostic agents. Agents that selectively bind to the foregoing polypeptides and genes also are provided.

Description

RELATED APPLICATIONS [0001] This application is a division of U.S. patent application Ser. No. 09 / 869486, filed Dec. 20, 1999, which is a national stage application under 35 U.S.C. §371 of International application PCT / US99 / 29996 designating the United States of America, filed Dec. 20, 1999, which claims the benefit under 35 U.S.C. §119(e) of U.S. provisional application 60 / 114,220, filed Dec. 30, 1998; of U.S. provisional application 60 / 120,018, filed Jan. 29, 1999; and of U.S. provisional application 60 / 140,415, filed Jun. 22, 1999.FIELD OF THE INVENTION [0002] This invention relates to nucleic acids coding for a novel family of calcium channel polypeptides, the encoded polypeptides, unique fragments of the foregoing, and methods of making and using same. BACKGROUND OF THE INVENTION [0003] Calcium channels are membrane-spanning, multi-subunit proteins that facilitate the controlled transport (“flux”) of Ca2+ ions into and out of cells. Cells throughout the animal kingdom, and at l...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/395C07K16/44
CPCC07K14/705A61K38/17
Inventor SCHARENBERG, ANDREW
Owner BETH ISRAEL DEACONESS MEDICAL CENT INC