The low-density lipoprotein class A module of the relaxin receptor (leucine-rich repeat containing G-protein coupled receptor 7): its role in signaling and trafficking to the cell membrane.
The relaxin receptor (LGR7, relaxin family peptide receptor 1) is a member of the leucine-rich repeat containing G protein-coupled receptors subgroup C. This and the LGR8 (relaxin family peptide receptor 2) receptor are unique in having a low-density lipoprotein class A (LDL-A) module at their N termini. This study was designed to show the role of the LDL-A in LGR7 expression and function. Point mutants for the conserved cysteines (Cys(47) and Cys(53)) and for calcium binding asparagine (Asp(58)), a mutant with deleted LDL-A domain and chimeric LGR7 receptor with LGR8 LDL-A all showed no cAMP response to human relaxins H1 or H2. We have shown that their cell surface delivery was uncompromised. The mutation of the putative N-linked glycosylation site (Asn(36)) decreased cAMP production and reduced cell surface expression to 37% of the wild-type LGR7. All point mutant, chimeric, and wild-type receptor proteins were expressed as the two forms. The immature or precursor form of the receptor was 80 kDa, whereas the mature receptor, delivered to the cell surface was 95 kDa. The glycosylation mutant was also expressed as two forms with appropriately smaller molecular masses. Deletion of the LDL-A module resulted in expression of the mature receptor only. These data suggest that the LDL-A module of LGR7 influences receptor maturation, cell surface expression, and relaxin-activated signal transduction.
Kern
András
A
Pacific Biosciences Research Center, University of Hawaii, 651 Ilalo Street, Honolulu, HI 96813, USA. kerna@pbrc.hawaii.edu
Agoulnik
Alexander I
AI
Bryant-Greenwood
Gillian D
GD
D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D017353:Gene Deletion; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006801:Humans; D026502:LDL-Receptor Related Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism* / Q000502:physiology*; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D021381:Protein Transport; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry* / Q000378:metabolism* / Q000502:physiology*; D018000:Receptors, Peptide; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction
D026502:LDL-Receptor Related Proteins; D008565:Membrane Proteins; D050505:Mutant Proteins; C417543:RXFP1 protein, human; D043562:Receptors, G-Protein-Coupled; D018000:Receptors, Peptide; D006026:Glycoside Hydrolases