Identification of an 11-kDa FKBP12-rapamycin-binding domain within the 289-kDa FKBP12-rapamycin-associated protein and characterization of a critical serine residue.

Chen, J; Zheng, X F; Brown, E J; Schreiber, S L

doi:10.1073/pnas.92.11.4947

Cited by 556PI3K/AKT/mTOR signaling in cancerSignaling Pathways in DiseaseProtein Kinase Regulation and GTPase Signaling

Identification of an 11-kDa FKBP12-rapamycin-binding domain within the 289-kDa FKBP12-rapamycin-associated protein and characterization of a critical serine residue.

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Chen, J, Zheng, X F, Brown, E J, Schreiber, S L·1995·Proceedings of the National Academy of Sciences

DOI 10.1073/pnas.92.11.4947PMC PMC41824

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Abstract

Complexed with its intracellular receptor, FKBP12, the natural product rapamycin inhibits G1 progression of the cell cycle in a variety of mammalian cell lines and in the yeast Saccharomyces cerevisae. Previously, a mammalian protein that directly associates with FKBP12-rapamycin has been identified and its encoding gene has been cloned from both human (designated FRAP) [Brown, E.J., Albers, M.W., Shin, T.B., Ichikawa, K., Keith, C.T., Lane, W.S. & Schreiber, S.L. (1994) Nature (London) 369, 756-758] and rat (designated RAFT) [Sabatini, D.M., Erdjument-Bromage, H., Lui, M., Tempst, P. & Snyder, S.H. (1994) Cell 78, 35-43]. The full-length FRAP is a 289-kDa protein containing a putative phosphatidylinositol kinase domain. Using an in vitro transcription/translation assay method coupled with proteolysis studies, we have identified an 11-kDa FKBP12-rapamycin-binding domain within FRAP. This minimal binding domain lies N-terminal to the kinase domain and spans residues 2025-2114. In addition, we have carried out mutagenesis studies to investigate the role of Ser2035, a potential phosphorylation site for protein kinase C within this domain. We now show that the FRAP Ser2035-->Ala mutant displays similar binding affinity when compared with the wild-type protein, whereas all other mutations at this site, including mimics of phosphoserine, abolish binding, presumably due to either unfavorable steric interactions or induced conformational changes.

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From Signal integration in the (m)TORC1 growth pathway (2018) · cites this paper as Chen et al., 1995

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Authors: Chen, J., Zheng, X. F., Brown, E. J., & Schreiber, S. L.
Affiliations: Howard Hughes Medical Institute (US)
ORCID: Chen, J
Zheng, X F
Brown, E J
Schreiber, S L
Venue: Proceedings of the National Academy of Sciences
Publisher: National Academy of Sciences
Published: 1995-05-23
Volume: 92
Issue: 11
Pages: 4947-4951
ISSN: 0027-8424 (ISSN-L)
1091-6490
DOI: 10.1073/pnas.92.11.4947
PMC: PMC41824
OpenAlex: W1987005832
Cited by: 556
References: 30
Citations by year: 2026: 9 · 2025: 7 · 2024: 22 · 2023: 19 · 2022: 15 · 2021: 19 · 2020: 11 · 2019: 20 · 2018: 17 · 2017: 26 · 2016: 18 · 2015: 29 · 2014: 19 · 2013: 27 · 2012: 30
FWCI: 3.60
Type: journal article
Language: en
Open access: green
www.ncbi.nlm.nih.gov
Locations: Proceedings of the National Academy of Sciences (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Funders: Howard Hughes Medical Institute
Damon Runyon Cancer Research Foundation
Harvard University
National Institutes of Health
National Institute of General Medical Sciences (GM38627)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › PI3K/AKT/mTOR signaling in cancer
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Signaling Pathways in Disease
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
Keywords: FRAP, FKBP12, rapamycin, protein kinase C phosphorylation sites, protein phosphorylation, protein conformational change, G1 cell cycle arrest
MeSH terms: Amino Acid Sequence, Animals, Binding Sites, Carrier Proteins, Circular Dichroism, Cloning, Molecular, DNA-Binding Proteins, Heat-Shock Proteins, Humans, Immunosuppressive Agents, Kinetics, Mammals, Molecular Sequence Data, Molecular Weight, Peptide Fragments, Polyenes, Protein Conformation, Recombinant Fusion Proteins, Saccharomyces cerevisiae, Serine, Transcription, Genetic, Protein Biosynthesis, Mutagenesis, Site-Directed, Tacrolimus, Sirolimus, Tacrolimus Binding Proteins, Rats

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Identification of an 11-kDa FKBP12-rapamycin-binding domain within the 289-kDa FKBP12-rapamycin-associated protein and characterization of a critical serine residue.

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