Hot Spot Mutagenesis Improves the Functional Expression of Unique Mammalian Odorant Receptors

Vertebrate animals detect odors through olfactory receptors (ORs), members of the G protein-coupled receptor (GPCR) family. Due to the difficulty in the heterologous expression of ORs, studies of their odor molecule recognition mechanisms have progressed poorly. Functional expression of most ORs in...

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Bibliographic Details
Published in:International journal of molecular sciences 2021-12, Vol.23 (1), p.277
Main Authors: Fukutani, Yosuke, Nakamura, Yuko, Muto, Nonoko, Miyanaga, Shunta, Kanemaki, Reina, Ikegami, Kentaro, Noguchi, Keiichi, Ohsawa, Ikuroh, Matsunami, Hiroaki, Yohda, Masafumi
Format: Article
Language:English
Subjects:
Amino acid substitution
Amino acids
Amino Acids - genetics
Animals
Binding sites
G protein-coupled receptors
Gene Expression Regulation
HEK293 Cells
Humans
Ligands
Loss of Function Mutation - genetics
Mammals - genetics
Membranes
Mice
Mutagenesis
Mutagenesis - genetics
Mutant Proteins - metabolism
Mutation
Mutation - genetics
Odor recognition
Odorant receptors
Odors
Protein transport
Proteins
Receptors, Odorant - agonists
Receptors, Odorant - chemistry
Receptors, Odorant - genetics
Residues
Sodium
Transmembrane domains
Vertebrates
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Summary:Vertebrate animals detect odors through olfactory receptors (ORs), members of the G protein-coupled receptor (GPCR) family. Due to the difficulty in the heterologous expression of ORs, studies of their odor molecule recognition mechanisms have progressed poorly. Functional expression of most ORs in heterologous cells requires the co-expression of their chaperone proteins, receptor transporting proteins (RTPs). Yet, some ORs were found to be functionally expressed without the support of RTP (RTP-independent ORs). In this study, we investigated whether amino acid residues highly conserved among RTP-independent ORs improve the functional expression of ORs in heterologous cells. We found that a single amino acid substitution at one of two sites (N 3.39 and 3.43) in their conserved residues (E and L, respectively) significantly improved the functional expression of ORs in heterologous cells. E and L also enhanced the membrane expression of RTP-dependent ORs in the absence of RTP. These changes did not alter the odorant responsiveness of the tested ORs. Our results showed that specific sites within transmembrane domains regulate the membrane expression of some ORs.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms23010277