Human substance P receptor (NK-1): Organization of the gene, chromosome localization, and functional expression of cDNA clones

Journal Article · Tue Nov 05 04:00:00 EST 1991 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00108a006· OSTI ID:5488092

Gerard, N P; Paquet, J L ^[1]; Garraway, L A ^[2]; Eddy, Jr, R L; Shows, T B ^[3]; Iijima, Hideya ^[4]; Gerard, C ^[5]

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1. Beth Israel Hospital, Boston, MA (United States) Children's Hospital, Boston, MA (United States) Harvard Medical School, Boston, MA (United States)
2. Children's Hospital, Boston, MA (United States) Harvard Medical School, Boston, MA (United States)
3. Roswell Park Memorial Inst., Buffalo, NY (United States)
4. Children's Hospital, Boston, MA (United States) Harvard School of Public Health, Boston, MA (United States)
5. Children's Hospital, Boston, MA (United States)

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{prime} and 3{prime} 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 {sup 125}I-BHSP with a K{sub d} of 0.35 {plus minus} 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.

• OSTI ID: 5488092
• Journal Information: Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 30:44; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

550401* -- Genetics-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
BETA DECAY RADIOISOTOPES
CHROMOSOMES
CLONING
DAYS LIVING RADIOISOTOPES
DNA
DNA HYBRIDIZATION
DNA POLYMERASES
DNA SEQUENCING
DNA-CLONING
ELECTRON CAPTURE RADIOISOTOPES
ENZYMES
GENE REGULATION
GENES
GENETIC MAPPING
HETEROCHROMOSOMES
HYBRIDIZATION
INTERMEDIATE MASS NUCLEI
INTERNAL CONVERSION RADIOISOTOPES
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KININS
MAPPING
MEMBRANE PROTEINS
MOLECULAR STRUCTURE
NUCLEI
NUCLEIC ACIDS
NUCLEOTIDYLTRANSFERASES
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PEPTIDES
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
POLYPEPTIDES
PROTEINS
RADIOISOTOPES
RADIORECEPTOR ASSAY
RECEPTORS
RECOMBINANT DNA
STRUCTURAL CHEMICAL ANALYSIS
TRACER TECHNIQUES
TRANSFERASES