Detection of lysozyme and alpha 2-macroglobulin--lysozyme complexes by immunoblotting.
An immunoblotting technique was developed to detect human lysozyme and lysozyme complexes in body fluids. The unoccupied binding capacity of proteins was demonstrated by addition of surplus lysozyme. The sensitivity of immunoblotting to the free enzyme in human albumin solution was less than 5 ng. In serum and pleural fluid, part of exogenous lysozyme was bound to alpha 2-macroglobulin (alpha 2-M). At high concentrations of lysozyme in leukemic sera, part of the enzyme formed an endogenous alpha 2-M complex. On the other hand, the formation of alpha 2-M complexes with exogenous lysozyme was especially striking in sera from nephrotic patients with elevated alpha 2-M. The findings corroborate with previous reports on lysozyme binding to purified alpha 2-M in vitro and suggest that the binding is concentration-dependent with respect to both reaction partners. In vivo the mechanism may provide a pathway for extrarenal lysozyme catabolism medicated by reticuloendothelial cells. No other binding proteins were seen in the present study: lysozyme did not bind to serum immunoglobulins in 35 samples with an immunoglobulin paraprotein, three samples with polyclonally elevated gamma-globulins, 20 other patient sera and 10 normal sera. Neither did lysozyme bind to urinary proteins in five samples from patients with myeloic leukemias nor in 10 samples from myeloma patients with urinary excretion of a monoclonal immunoglobulin light chain.
Mörsky
P
P
Department of Clinical Chemistry, Tampere University Central Hospital, Finland.
D006801:Humans; D015151:Immunoblotting; D007153:Immunologic Deficiency Syndromes / Q000097:blood / Q000652:urine; D007951:Leukemia, Myeloid / Q000097:blood / Q000652:urine; D009113:Muramidase / Q000032:analysis* / Q000097:blood / Q000652:urine; D009401:Nephrosis / Q000097:blood; D010994:Pleura / Q000378:metabolism; D010998:Pleurisy / Q000097:blood / Q000652:urine; D000511:alpha-Macroglobulins / Q000032:analysis* / Q000097:blood / Q000652:urine
D000511:alpha-Macroglobulins; D009113:Muramidase