Cloning, sequencing, and functional characterization of the murine 46-kDa mannose 6-phosphate receptor.
We have cloned and sequenced the 2175-nucleotide, full-length cDNA for the mouse 46-kDa Man 6-P receptor (46MPR) and studied its functional properties in stably transfected mouse L cells which do not express the insulin-like growth factor-II receptor/mannose 6-phosphate receptor (IGF-IIR/MPR). The 278-amino acid sequence deduced from the cDNA for the murine 46MPR shows 19 amino acid differences from that of the human 46MPR, none of which are found in the 68-amino acid cytoplasmic tail. Binding of ligand to the murine 46MPR in permeabilized cells showed a pH optimum of 6.5, was completely inhibited by Man 6-P, and was stimulated by divalent cations. Mn2+ was more effective than Ca2+ or Mg2+. Endocytosis was demonstrated at pH 6.5 and was stimulated 4-7-fold by Mn2+. In its responsiveness to divalent cations and its preference for Mn2+, the murine 46MPR resembled the bovine 46MPR more than the human 46MPR. It was even less efficient than the human receptor in its ability to mediate endocytosis in transfected murine cells. It was also no more efficient than the human 46MPR in correcting the sorting defect of IGF-IIR/MPR-deficient mouse L cells. We conclude that the previously observed relative inefficiency of the human 46MPR in sorting enzymes to lysosomes in murine cells is a property of the 46MPR itself and not a manifestation of studying its expression in a heterologous cell line.
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Edward A. Doisy Department of Biochemistry and Molecular Biology, St. Louis University School of Medicine, Missouri 63104.
Grubb
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D000818:Animals; D001483:Base Sequence; D002413:Cations, Divalent; D002460:Cell Line; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004705:Endocytosis; D005966:Glucuronidase / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008360:Mannosephosphates / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D017527:Receptor, IGF Type 2; D011956:Receptors, Cell Surface / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection
D002413:Cations, Divalent; D008360:Mannosephosphates; D017527:Receptor, IGF Type 2; D011956:Receptors, Cell Surface; D005966:Glucuronidase