Distinct physicochemical characteristics of human mannose binding protein expressed by individuals of differing genotype.
Mannose binding protein (MBP) is a serum collectin (collagenous lectin) believed to be of importance in innate immunity. Three point mutations, in codons 52, 54 and 57 of exon 1 of the human MBP gene, have been predicted to affect the tertiary structure of the collagenous region of the protein, and are known to be associated with low serum concentrations of MBP. However, other groups working with recombinant mutant proteins have claimed that the proteins are expressed and assembled normally. The aim of the present investigation was to characterize the effects of these mutations on the physicochemical nature of MBP that is present in the circulation in vivo, and for this we used polyacrylamide gel electrophoresis, gel filtration and sucrose density gradient centrifugation, followed by immunoblotting and enhanced chemiluminescence. The circulating wild-type MBP appeared to comprise a mixture of polymers formed from two to eight subunits (each based on three identical 32,000 MW polypeptide chains) of apparent molecular weights 200,000-700,000, with dimers and trimers constituting the predominant forms. Individuals homozygous for the codon 54 or 57 mutation had dramatically reduced concentrations of serum MBP, mainly comprising material of an apparent molecular weight of 120,000-130,000. Heterozygous individuals showed characteristics of both phenotypes. In contrast to the results obtained with artificial expression systems, our data suggest that individuals homozygous for the MBP mutations have very little circulating protein and that this comprises mainly low molecular weight material.
Lipscombe
R J
RJ
Molecular Immunology Unit, St Mary's Hospital Medical School, Imperial College of Science and Technology, London, UK.
Sumiya
M
M
Summerfield
J A
JA
Turner
M W
MW
D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics*; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005838:Genotype; D006801:Humans; D015151:Immunoblotting; D008358:Mannose / Q000378:metabolism*; D037241:Mannose-Binding Lectins; D008970:Molecular Weight; D017354:Point Mutation
D002352:Carrier Proteins; D037241:Mannose-Binding Lectins; D008358:Mannose