title: Human substance P receptor (NK-1): organization of the gene, chromosome localization, and functional expression of cDNA clones.
abstract: The gene for the human substance P receptor (NK-1) was cloned using cDNA probes made by the polymerase chain reaction from primers based on the rat sequence. The gene spans 45-60 kb and is contained in five exons, with introns interrupting at sites homologous to those in the NK-2 receptor gene. Analysis of restriction digests of genomic DNA from mouse/human cell hybrids indicates the NK-1 receptor is a single-copy gene located on human chromosome 2. Polymerase chain reaction using primers based on the 5' and 3' ends of the coding sequence was used to generate full-length cDNAs from human lung and from IM9 lymphoblast cells. When transfected into COS-7 cells, the NK-1 receptor binds 125I-BHSP with a Kd of 0.35 +/- 0.07 nM and mediates substance P induced phosphatidylinositol metabolism. The receptor is selective for substance P; the relative affinity for neurokinin A and neurokinin B is 100- and 500-fold lower, respectively. Human IM9 lymphoblast cells express relatively high levels of the NK-1 receptor, and Northern blot analysis indicates modulation of mRNA levels by glucocorticoids and growth factors, suggesting that this cell line may be useful as a model for studying the control of NK-1 receptor gene expression.
authors.affiliation: Department of Medicine, Beth Israel Hospital, Boston, Massachusetts 02215.
authors.forename: N P
authors.initials: NP
authors.lastname: Gerard
authors.forename: L A
authors.initials: LA
authors.lastname: Garraway
authors.forename: R L
authors.initials: RL
authors.lastname: Eddy
authors.forename: T B
authors.initials: TB
authors.lastname: Shows
authors.forename: H
authors.initials: H
authors.lastname: Iijima
authors.forename: J L
authors.initials: JL
authors.lastname: Paquet
authors.forename: C
authors.initials: C
authors.lastname: Gerard
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D002874:Chromosome Mapping; D002889:Chromosomes, Human, Pair 2; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D005091:Exons; D015870:Gene Expression; D005786:Gene Expression Regulation; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D018040:Receptors, Neurokinin-1; D017981:Receptors, Neurotransmitter / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: D018040:Receptors, Neurokinin-1; D017981:Receptors, Neurotransmitter; D004247:DNA