Cloning and expression of the human substance K receptor and analysis of its role in mitogenesis.
The primary structure of the human substance K receptor was established from the sequences of complementary DNA clones isolated from a human jejunal complementary DNA library. It consists of 398 amino acids, including seven putative transmembrane regions. The gene for the human substance K receptor was localized to chromosome region 10p13-10q23, a region with frequent chromosomal abnormalities. The human substance K receptor was expressed in transfected NIH-3T3 cells lacking endogenous substance K receptors, and Scatchard analysis of 125I-labeled substance K binding indicates approximately 100,000 receptors/cell with a single dissociation constant of 12 nM. Covalent cross-linking experiments utilizing 125I-substance K and three different chemical cross-linking reagents (disuccinimidyl suberate, disuccinimidyl tartrate, or 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide-HCl) demonstrate an apparent molecular weight of 45,000, consistent with little or no N-linked glycosylation. The binding of substance K to its receptor on transfected cells led to a rapid increase in the production of total inositol phosphates and the release of Ca2+ from internal stores. Growth of the cells transfected with the human substance K receptor is stimulated by the addition of substance K to the medium to a level similar to 10% serum. Therefore, the human substance K receptor can function as a growth factor receptor when expressed in mouse 3T3 cells.
Kris
R M
RM
Department of Pharmacology, New York University School of Medicine, New York 10016.
South
V
V
Saltzman
A
A
Felder
S
S
Ricca
G A
GA
Jaye
M
M
Huebner
K
K
Kagan
J
J
Croce
C M
CM
Schlessinger
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D005347:Fibroblasts / Q000378:metabolism; D006133:Growth Substances / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D015288:Neurokinin A / Q000502:physiology*; D018041:Receptors, Neurokinin-2; D017981:Receptors, Neurotransmitter / Q000096:biosynthesis / Q000737:chemistry / Q000502:physiology*; D012689:Sequence Homology, Nucleic Acid; D015398:Signal Transduction / Q000502:physiology
D006133:Growth Substances; D018041:Receptors, Neurokinin-2; D017981:Receptors, Neurotransmitter; D015288:Neurokinin A