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Murine histidine-rich glycoprotein: cloning, characterization and cellular origin
1Division of Immunology and Cell Biology, The John Curtin School of Medical Research, Australian National University, Canberra, Australian Capital Territory, Australia.
Immunology and Cell Biology
|June 10, 2000
Summary
Histidine-rich glycoprotein (HRG) is primarily synthesized in the liver, not immune cells. This study clarifies HRG gene expression and localization in mice and rats, debunking immune cell synthesis theories.
Area of Science:
- Biochemistry
- Genetics
- Immunology
Background:
- Histidine-rich glycoprotein (HRG) is a vertebrate plasma protein involved in immune response and blood clotting.
- Previous research has suggested, controversially, that immune cells like monocytes and megakaryocytes synthesize HRG.
Purpose of the Study:
- To isolate and determine the cDNA sequences for mouse and rat HRG.
- To investigate the expression localization of HRG mRNA in mice.
- To clarify the cellular origin of HRG in the immune system.
Main Methods:
- cDNA sequencing
- Northern blot analysis
- Reverse transcriptase-polymerase chain reaction (RT-PCR)
- Genomic Southern blot analysis
Main Results:
- Mouse and rat HRG cDNA sequences were determined, revealing a conserved three-domain structure and high amino acid identity with human HRG.
- Mouse HRG mRNA (1.7 kb) was localized specifically to the liver.
- RT-PCR analysis detected no HRG mRNA in mouse immune tissues (spleen, thymus, lymph node, bone marrow, peripheral blood leucocytes).
- Genomic Southern blot indicated HRG is a single-copy gene in mice.
Conclusions:
- HRG is predominantly synthesized in the liver.
- Immune cell expression of HRG is likely due to the uptake of circulating plasma HRG, not de novo synthesis.
- The HRG gene is a single-copy gene in the mouse genome.
