Discrimination between alternatively spliced STP-A and -B isoforms of CD46.
CD46 (membrane cofactor protein; MCP) is ubiquitously expressed on nucleated human cells; it has a protective function, binding C3b and C4b, which are then cleaved by serum factor I. CD46 molecules (55,000-65,000 MW) have four short consensus repeats (SCR): the function of SCR-1 and -2 is unknown; SCR-3 and -4 bind C3b and C4b. These are succeeded by the STP region, which can contain three separate regions (STP-A, -B, -C) rich in serine, threonine and proline and which are heavily glycosylated, succeeded by transmembrane and cytoplasmic tail regions (of which there are several). Multiple isoforms exist due to the different splicing of exons: STP-A and -B can thus be present or absent. So far these products can only be detected separately by polymerase chain reaction (PCR) and RNA studies; we now describe their detection by anti-peptide antibodies. Peptides whose sequences corresponded with those of STP-A and STP-B were synthesized and used for the immunization of mice; although they differ in only seven of 21 amino acids, monoclonal antibodies (mAb) that reacted specifically with STP-A but not with STP-B, and mAb that reacted specifically with STP-B but not with STP-A, were produced; these reacted specifically with native CD46 on human tissues and cell lines. STP-A mAb reacted with tissues in which STP-A RNA had been found, some leukaemias and cell lines; in normal tissue expression was mainly found in the intestine (large and small) and salivary gland. Anti-STP-B reacted with most tissues and cell lines. The antibodies should be of use in defining the expression and function of CD46 in different tissues.
Xing
P X
PX
Austin Research Institute, Austin Hospital, Heidelberg, Victoria, Australia.
Russell
S
S
Prenzoska
J
J
McKenzie
J F
JF
D000116:Acetylgalactosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D015703:Antigens, CD / Q000737:chemistry / Q000276:immunology*; D001593:Benzyl Compounds / Q000494:pharmacology; D015153:Blotting, Western; D002469:Cell Separation / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D007124:Immunoenzyme Techniques; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000276:immunology*; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000276:immunology; D010446:Peptide Fragments / Q000276:immunology
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D001593:Benzyl Compounds; C497857:CD46 protein, human; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; C080739:benzyl-alpha-N-acetylgalactosamine; D000116:Acetylgalactosamine