In vitro phosphorylation of the 46-kDa mannose 6-phosphate receptor by casein kinase II. Structural requirements for efficient phosphorylation.
Some steps in the receptor-mediated transport of newly synthesized mannose 6-phosphate-containing lysosomal enzymes are assumed to be accompanied by changes in the phosphorylation state of receptors. In vitro, the metabolically phosphorylated 46-kDa mannose 6-phosphate receptor (MPR 46) was dephosphorylated by protein phosphatase 2A. The synthetic cytoplasmic domain of MPR 46 was phosphorylated in vitro by casein kinase II. Tryptic phosphopeptide mapping showed that casein kinase II phosphorylates MPR 46 in vitro at the same site that is phosphorylated in vivo. Inhibition studies using synthetic peptides corresponding to different amino acid sequences of the cytoplasmic tail of MPR 46 revealed that the sequence 26-32 (ADGCDFV) contribute to efficient phosphorylation of serine 56. Baby hamster kidney cells were transfected with wild type human MPR 46 cDNA or cDNAs containing mutations in the cytoplasmic tail and assayed for their phosphorylation state in vivo. The phosphorylation of mutant receptors with deleted residues 23-28 (NLVADG) was strongly reduced. These data indicate that residues on the N-terminal side of the phosphorylatable serine 56 may influence the efficiency with which a casein-kinase II-like kinase phosphorylates MPR 46.
Körner
C
C
Institute of Biochemistry II, University of Göttingen, Federal Republic of Germany.
Herzog
A
A
Weber
B
B
Rosorius
O
O
Hemer
F
F
Schmidt
B
B
Braulke
T
T
D000595:Amino Acid Sequence; D000818:Animals; D047390:Casein Kinase II; D002460:Cell Line; D006224:Cricetinae; D006801:Humans; D007668:Kidney; D007700:Kinetics; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D010449:Peptide Mapping; D010455:Peptides / Q000494:pharmacology; D010748:Phosphopeptides / Q000737:chemistry / Q000302:isolation & purification; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D010766:Phosphorylation; D054648:Protein Phosphatase 2; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D017527:Receptor, IGF Type 2 / Q000302:isolation & purification / Q000378:metabolism*; D014162:Transfection
D010446:Peptide Fragments; D010455:Peptides; D010748:Phosphopeptides; D017527:Receptor, IGF Type 2; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2