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Mouse K-Cl cotransporter KCC1: cloning, mapping, pathological expression, and functional regulation.
W Su1, B E Shmukler, M N Chernova
1Molecular Medicine and Renal Units, Beth Israel Deaconess Medical Center, Boston 02215, USA.
The American Journal of Physiology
|November 24, 1999
Summary
Researchers identified the K-Cl cotransporter 1 (KCC1) protein in red blood cells, finding its abundance increases with red blood cell production and is linked to certain blood disorders.
Area of Science:
- Molecular Biology
- Physiology
- Genetics
Background:
- K-Cl cotransporter (KCC1) mRNA is widespread, but KCC1 protein and activity are poorly characterized.
- Endogenous KCC1 polypeptide detection has been lacking, hindering functional studies.
Purpose of the Study:
- To clone and characterize the mouse erythroid KCC1 (mKCC1) cDNA and protein.
- To investigate the expression, distribution, and regulation of KCC1 in erythroid cells.
Main Methods:
- Cloning of mKCC1 cDNA and genomic regions.
- Generation of anti-peptide antibodies for immunoblot and immunoprecipitation.
- Analysis of mKCC1 mRNA and protein expression in various tissues and conditions.
- Functional studies of mKCC1 in Xenopus oocytes.
Main Results:
- mKCC1 cDNA and gene were cloned and mapped.
- Specific antibodies detected widespread mKCC1 mRNA and protein expression.
- Erythroid KCC1 polypeptide levels correlated with reticulocyte counts in normal and disease states.
- Functional assays confirmed KCC1 activity and erythroid-specific regulation.
Conclusions:
- KCC1 protein is present in erythrocytes and its abundance is modulated by erythropoiesis.
- The developed reagents and findings facilitate KCC1 research and understanding of related diseases.