Structural Determinants of the Insulin Receptor-related Receptor Activation by Alkali

Deyev, I. E.; Mitrofanova, Alla; Zhevlenev, Egor S.; Radionov, N. V.; Berchatova, Anastasiya A.; Popova, N. V.; Serova, O. V.; Petrenko, Alexander G.

doi:10.1074/jbc.m113.483172

Cited by 24 publications

(27 citation statements)

References 28 publications

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“…A dose-dependent and time course effect of TM peptides on receptor phosphorylation was detected by Western blot. To detect the total protein as a loading control for phosphorylated receptors, the membrane was stripped with 0.2 M glycine and 1% SDS for 15 min, blocked, and incubated with anti-TrkA, TrkB, TrkC, and FGFR1 antibodies (52,53).…”

Section: In Vitro Phosphorylation Assaymentioning

confidence: 99%

The transmembrane domain of the p75 neurotrophin receptor stimulates phosphorylation of the TrkB tyrosine kinase receptor

Saadipour

MacLean

Pirkle

et al. 2017

Journal of Biological Chemistry

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The function of protein products generated from intramembraneous cleavage by the γ-secretase complex is not well defined. The γ-secretase complex is responsible for the cleavage of several transmembrane proteins, most notably the amyloid precursor protein that results in Aβ, a transmembrane (TM) peptide. Another protein that undergoes very similar γ-secretase cleavage is the p75 neurotrophin receptor. However, the fate of the cleaved p75 TM domain is unknown. p75 neurotrophin receptor is highly expressed during early neuronal development and regulates survival and process formation of neurons. Here, we report that the p75 TM can stimulate the phosphorylation of TrkB (tyrosine kinase receptor B). phosphorylation experiments indicated that a peptide representing p75 TM increases TrkB phosphorylation in a dose- and time-dependent manner. Moreover, mutagenesis analyses revealed that a valine residue at position 264 in the rat p75 neurotrophin receptor is necessary for the ability of p75 TM to induce TrkB phosphorylation. Because this residue is just before the γ-secretase cleavage site, we then investigated whether the p75(αγ) peptide, which is a product of both α- and γ-cleavage events, could also induce TrkB phosphorylation. Experiments using TM domains from other receptors, EGFR and FGFR1, failed to stimulate TrkB phosphorylation. Co-immunoprecipitation and biochemical fractionation data suggested that p75 TM stimulates TrkB phosphorylation at the cell membrane. Altogether, our results suggest that TrkB activation by p75(αγ) peptide may be enhanced in situations where the levels of the p75 receptor are increased, such as during brain injury, Alzheimer's disease, and epilepsy.

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Section: In Vitro Phosphorylation Assaymentioning

confidence: 99%

The transmembrane domain of the p75 neurotrophin receptor stimulates phosphorylation of the TrkB tyrosine kinase receptor

Saadipour

MacLean

Pirkle

et al. 2017

Journal of Biological Chemistry

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“…In vivo experiments using mice with IRR gene knockout demonstrated that this receptor is involved in the regulation of renal excretion of excess alkali in the form of bicarbonate [ 6 , 7 ]. Mapping of the regions determining the pH sensitivity of IRR has shown that several extracellular domains are responsible for receptor activation [ 5 , 8 ], which ensures positive cooperation in activation (the Hill’s coefficient being ~ 2.4) [ 5 , 9 ].…”

Section: Introductionmentioning

confidence: 99%

Determination of Alkali-Sensing Parts of the Insulin Receptor-Related Receptor Using the Bioinformatic Approach

2015

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IRR (insulin receptor-related receptor) is a receptor tyrosine kinase belonging to the insulin receptor family, which also includes insulin receptor and IGF-IR receptor. We have previously shown that IRR is activated by extracellular fluid with pH > 7.9 and regulates excess alkali excretion in the body. We performed a bioinformatic analysis of the pH-sensitive potential of all three members of the insulin receptor family of various animal species (from frog to man) and their chimeras with swapping of different domains in the extracellular region. An analysis using the AcalPred program showed that insulin receptor family proteins are divided into two classes: one class with the optimal working pH in the acidic medium (virtually all insulin receptor and insulin-like growth factor receptor orthologs, except for the IGF-IR ortholog from Xenopus laevis) and the second class with the optimal working pH in the alkaline medium (all IRR orthologs). The program had predicted that the most noticeable effect on the pH-sensitive property of IRR would be caused by the replacement of the L1 and C domains in its extracellular region, as well as the replacement of the second and third fibronectin repeats. It had also been assumed that replacement of the L2 domain would have the least significant effect on the alkaline sensitivity of IRR. To test the in silico predictions, we obtained three constructs with swapping of the L1C domains, the third L2 domain, and all three domains L1CL2 of IRR with similar domains of the insulin-like growth factor receptor. We found that replacement of the L1C and L1CL2 domains reduces the receptor’s ability to be activated with alkaline pH, thus increasing the half-maximal effective concentration by about 100%. Replacement of the L2 domain increased the half-maximal effective concentration by 40%. Thus, our results indicate the high predictive potential of the AcalPred algorithm, not only for the pH-sensitive enzymes, but also for pH-sensitive receptors.

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“…The IR family receptors are structurally unique when compared to other RTKs; fully processed IR, IGF1R, and IRR are disulfide-linked preformed dimers, as opposed to monomers that dimerize upon activation [2–5]. IRR is the least understood within the IR family and has no known ligand, although IRR may act as an extracellular alkali sensor [6]. On the other hand, IR and IGF1R have been extensively studied because of their importance in the regulation of many normal cellular functions, and because of their roles in diseases such as diabetes and cancer [7,8].…”

Section: Introductionmentioning

confidence: 99%

Expression and purification of functional insulin and insulin-like growth factor 1 holoreceptors from mammalian cells

Bovi

Miller

2017

Analytical Biochemistry

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The insulin receptor (IR) and insulin-like growth factor 1 receptor (IGF1R) are receptor tyrosine kinases (RTKs) involved in the regulation of many important cellular processes. The current proposed models of activation are derived from structural studies using soluble extracellular domains and cytoplasmic tyrosine kinase domains. Preparations of full length IR and IGF1R have been hampered by the need for unconventional affinity chromatography resins and/or harsh eluting conditions. Here, we present a purification protocol to obtain full-length, detergent solubilized IR and IGF1R at quantities suitable for biochemical and structural characterization. We screened a panel of 24 structurally diverse detergents for optimal ligand activation. The receptors purified in n-dodecyl-β-D-maltoside showed ligand-stimulated autophosphorylation and kinase activity, suggesting an intact transmembrane signaling mechanism. This convenient purification protocol can be used to produce high quantities of IR, IGF1R, or other RTKs, and can be adapted for other challenging membrane proteins.

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Structural Determinants of the Insulin Receptor-related Receptor Activation by Alkali

Cited by 24 publications

References 28 publications

The transmembrane domain of the p75 neurotrophin receptor stimulates phosphorylation of the TrkB tyrosine kinase receptor

The transmembrane domain of the p75 neurotrophin receptor stimulates phosphorylation of the TrkB tyrosine kinase receptor

Determination of Alkali-Sensing Parts of the Insulin Receptor-Related Receptor Using the Bioinformatic Approach

Expression and purification of functional insulin and insulin-like growth factor 1 holoreceptors from mammalian cells

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