Identification of the putative mannose 6-phosphate receptor (MPR 46) protein in the invertebrate mollusc.
Mannose 6-phosphate receptor (MPR 300) protein was earlier affinity purified on phosphomannan gel from the membrane extracts of whole animal acetone powder of a mollusc, unio, in the presence of EDTA (Udaya Lakshmi, Y., Radha, Y., Hille-Rehfeld, A., von Figura, K., and Siva Kumar, N. (1999) Biosci. Rep. 19:403-409). In the present study we demonstrate that the unio also contains the putative mannose 6-phosphate receptor (MPR 46) that can be purified on the same gel in presence of divalent metal ions (10 mM each of calcium, manganese, and magnesium), and in the absence of sodium chloride and at pH 6.5. Chicken and Fish cell MPR 46 proteins were purified under these conditions (Siva Kumar, N., Udaya Lakshmi, Y., Hille-Rehfeld, A., and von Figura, K. (1999) Comp. Biochem. & PhysioL 123B:261-265). The authenticity of the receptor is further confirmed by its ability to react with the MSC1 antibody that is specific for MPR 46 protein. Additional evidence for the presence of MPR 46 in molluscs could be obtained by metabolic labeling of mollusc cells Biomphalaria glabrata (Bg cells) with [35S] methionine and cysteine, and passing the labeled membrane extract on phosphomannan gel (at pH 6.5 and 7.0). On elution with mannose 6-phosphate, followed by immunoprecipitation of the column fractions, we identified the putative MPR 46 protein in the Bg cells. When Bg cell MPR 46 was deglycosylated along with chicken MPR 46 (control) both species yielded a single polypeptide corresponding to molecular mass of 26 kDa, suggesting that both contain the same receptor protein.
Nadimpalli
Siva Kumar
SK
Protein Biochemistry Laboratory, Department of Biochemistry, University of Hyderabad, Hyderabad 500 046, India. nsksl@uohyd.ernet.in
von Figura
Kurt
K
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity / Q000502:physiology; D001700:Biomphalaria / Q000166:cytology / Q000378:metabolism*; D002462:Cell Membrane / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D019143:Evolution, Molecular; D008360:Mannosephosphates / Q000378:metabolism*; D008970:Molecular Weight; D010802:Phylogeny; D017527:Receptor, IGF Type 2 / Q000235:genetics / Q000302:isolation & purification*; D013347:Subcellular Fractions / Q000737:chemistry; D014714:Vertebrates / Q000378:metabolism
D000906:Antibodies; D008360:Mannosephosphates; D017527:Receptor, IGF Type 2; C027693:mannose-6-phosphate