Mutational and secondary structural analysis of the basolateral sorting signal of the polymeric immunoglobulin receptor.
The 17-juxtamembrane cytoplasmic residues of the polymeric immunoglobulin receptor contain an autonomous basolateral targeting signal that does not mediate rapid endocytosis (Casanova, J. E., G. Apodaca, and K. E. Mostov. Cell. 66:65-75). Alanine-scanning mutagenesis identifies three residues in this region, His656, Arg657, and Val660, that are most essential for basolateral sorting and two residues, Arg655 and Tyr668, that play a lesser role in this process. Progressive truncations suggested that Ser664 and Ile665 might also play a role in basolateral sorting. However, mutation of these residues to Ala or internal deletions of these residues did not affect basolateral sorting, indicating that these residues are probably not required for basolateral sorting. Two-dimensional NMR spectroscopy of a peptide corresponding to the 17-mer signal indicates that the sequence Arg658-Asn-Val-Asp661 has a propensity to adopt a beta-turn in solution. Residues COOH-terminal to the beta-turn (Arg662 to Arg669) seem to take up a nascent helix structure in solution. Substitution of Val660 with Ala destabilizes the turn, while mutation of Arg657 to Ala does not appear to affect the turn structure. Neither mutation detectably altered the stability of the nascent helix in the COOH-terminal portion of the peptide.
Aroeti
B
B
Department of Anatomy, University of California, San Francisco 94143.
Kosen
P A
PA
Kuntz
I D
ID
Cohen
F E
FE
Mostov
K E
KE
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D002460:Cell Line; D004247:DNA; D004252:DNA Mutational Analysis; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry; D021382:Protein Sorting Signals / Q000737:chemistry* / Q000235:genetics; D017433:Protein Structure, Secondary; D011971:Receptors, Immunologic; D012635:Secretory Component / Q000737:chemistry* / Q000235:genetics; D013329:Structure-Activity Relationship
D009842:Oligopeptides; D021382:Protein Sorting Signals; D011971:Receptors, Immunologic; D012635:Secretory Component; D004247:DNA