Human leptin forms complexes with alpha 2-macroglobulin which are recognized by the alpha 2-macroglobulin receptor/low density lipoprotein receptor-related protein.
OBJECTIVE
To identify binding proteins of leptin in human plasma.


METHODS
Binding was evaluated by electrophoresis, size exclusion chromatography (SEC), Western blotting, and radioisotope labeling. Quantification of leptin and the different forms of alpha2-macroglobulin (alpha2-M) was performed by ELISA.


RESULTS
Leptin interacts with the proteinase inhibitor, alpha2-M. 125I-labeled leptin specifically binds to the transformed inhibitor, which arises by reaction with proteinases or with reactive primary amines. No leptin binding was observed to the native alpha2-M, which abundantly occurs in plasma. The complex formation between leptin and alpha2-M was found to proceed within minutes and was stable, as it resisted separation by SEC and electrophoresis. The Kd of the complex was 2.14 +/- 0.78 micromol/l. Complex formation with transformed alpha2-M did not interfere with the immunological determination of leptin in plasma. The leptin-alpha2-M complex was found to be recognized by the alpha2-M receptor/low density lipoprotein receptor-related protein. By computer analysis, a simple model is presented showing that the degree of transformation of alpha2-M may significantly influence the leptin concentration in blood.


CONCLUSIONS
The proteinase inhibitor, alpha2-M, may act as a leptin-binding protein in human plasma. Binding of leptin to transformed alpha2-M and its rapid clearance by the alpha2-M receptor may significantly influence the bioavailability of leptin in human plasma.
Institute of Biochemistry, University of Leipzig, Germany.
G
G
Birkenmeier
I
I
Kämpfer
J
J
Kratzsch
W
W
Schellenberger
D000328:Adult; D002352:Carrier Proteins / Q000097:blood; D005260:Female; D006801:Humans; D020738:Leptin; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D046911:Macromolecular Substances; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D011971:Receptors, Immunologic / Q000097:blood*; D011973:Receptors, LDL / Q000097:blood*; D054411:Receptors, Leptin; D000511:alpha-Macroglobulins / Q000378:metabolism*
D002352:Carrier Proteins; C515448:LEPR protein, human; D020738:Leptin; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D046911:Macromolecular Substances; D011506:Proteins; D011971:Receptors, Immunologic; D011973:Receptors, LDL; D054411:Receptors, Leptin; D000511:alpha-Macroglobulins