key: cord-0221738-pv2wvgbt authors: Maaroufi, Halim title: Interactions of SARS-CoV-2 spike protein and transient receptor potential (TRP) cation channels could explain smell, taste, and/or chemesthesis disorders date: 2021-01-15 journal: nan DOI: nan sha: 14f810f973cbf613579abc19bf1945f53c1fc549 doc_id: 221738 cord_uid: pv2wvgbt A significant subset of patients infected by SARS-CoV-2 presents olfactory, taste, and/or chemesthesis (OTC) disorders (OTCD). These patients recover rapidly, eliminating damage of sensory nerves. Discovering that S protein contains two ankyrin repeat binding motifs (S-ARBMs) and some TRP cation channels, implicated in OTC, have ankyrin repeat domains (TRPs-ARDs), I hypothesized that interaction of S-ARBMs and TRPs-ARDs could dysregulate the function of the latter and thus explains OTCD. Of note, some TRPs-ARDs are expressed in the olfactory epithelium, taste buds, trigeminal neurons in the oronasal cavity and vagal neurons in the trachea/lungs. Furthermore, this hypothesis is supported by studies that have shown: (i) respiratory viruses interact with TRPA1 and TRPV1 on sensory nerves and epithelial cells in the airways, (ii) the respiratory pathophysiology in COVID-19 patients is similar to lungs injuries produced by the sensitization of TRPV1 and TRPV4, and (iii) resolvin D1 and D2 shown to reduce SARS-CoV-2-induced inflammation, directly inhibit TRPA1, TRPV1, TRPV3 and TRPV4. Herein, results of blind dockings of S-ARBMs, 408-RQIAPG-413 (in RBD but distal from the ACE-2 binding region) and 905-RFNGIG-910 (in HR1), into TRPA1, TRPV1 and TRPV4 suggest that S-ARBMs interact with ankyrin repeat 6 of TRPA1 near an active site, and ankyrin repeat 3-4 of TRPV1 near cysteine 258 supposed to be implicated in the formation of inter-subunits disulfide bond. These findings suggest that S-ARBMs affect TRPA1, TRPV1 and TRPV4 function by interfering with channel assembly and trafficking. After an experimental confirmation of these interactions, among possible preventive treatments against COVID-19, the use of pharmacological manipulation (probably inhibition) of TRPs-ARDs to control or mitigate sustained pro-inflammatory response. that conveys chemesthesis, is a separate sensory system with distinct peripheral and central neural mechanisms (Shepherd, 2006; Green, 2012) . The trigeminal nerve may serve as a route for entry of pathogens into the brain, thus the RNA of coronavirus, mouse hepatitis virus strain JHM was detected in the trigeminal and olfactory nerves (Perlman et al., 1989) . 2006; Venkatachalam and Montell, 2007) . They are homo or hetero-tetramers, each subunit forms six transmembrane segments (S1-S6) and a pore-loop between S5 and S6 that corresponds to a voltage-sensor-like domain and a pore domain. Each TRP channel subfamily is characterized by a unique cytoplasmic domain . The cytoplasmic N-terminal sequence of TRPA1, TRPC1, 3-7 and TRPV1-6 To prevent the development of severe COVID-19 form, it is necessary to understand the cellular basis of SARS-CoV-2 infection. Identifying sensory-neural mechanisms responsible of OTCD could help to find treatments to stop or mitigate COVID-19 development. Here, the potential molecular mechanism responsible of OTCD in SARS-CoV-2 infected patients is presented. Discovering that S protein contains two S-ARBMs and some TRPs cation channels, implicated in OTC, have ARDs, I hypothesized that the interaction of S-ARBMs and TRPs-ARDs could dysregulate the function of TRP channels and thus explains OTCD. Blind docking results suggested that probably S-ARBMs affect TRPs-ARDs function by interfering with channel assembly and trafficking. The eukaryotic linear motif (ELM) resource (http://elm.eu.org/) revealed in SARS-CoV-2 S protein two short linear motifs (SLiMs) known as ARBMs (R-x-x- CoV, but absent in MERS-CoV (Fig. 1A, B) . Interestingly, figure 1C shows that SARS-CoV-2, SARS-CoV, bat RaTG13 and pangolin GX-P5L S proteins with ARBMs in both S1 and S2 subunits are phylogenetically close. Of note, S2 subunit of the other beta- ARBMs are present in S protein of SARS-CoV-2 and SARS-CoV (Fig. 1A, B) , and their interaction with TRPs-ARDs could explain OTCD. Intriguingly, only one single case of anosmia was reported during the SARS-CoV pandemic (Hwang, 2006) , against almost millions of cases in SARS-CoV-2 (von Bartheld et al., 2020). In the beginning of the SARS-CoV-2 pandemic, loss of olfactory and taste were considered as anecdotal. May be in the SARS-CoV pandemic, these first symptoms were also considered as anecdotal due to infection cases were not large (little more than 8,000) (Peiris et al., 2003) , against millions of cases in SARS-CoV-2, to reveal clearly that loss of olfactory and/or taste are linked to SARS-CoV infection. In addition, it has been observed that Caucasians had a 3- Scanning SARS-CoV-2 S protein sequence with DisEMBL software (Linding et al., 2003) revealed in the RBD a hot disordered loop, 404 RGDEVRQIAPGQTGKIA 419 , that contains four motifs: 408 RQIAPG 413 motif, reversed consensus pattern (TQ) phosphorylation (reading from C-to N-terminus) (Torshin, 2000) , phosphothreonine motif (T-x-x-I) and RGD motif (Table 1) . The Thr415 of the reversed consensus pattern (TQ) (reading from C-to N-terminus) in 404 RGDEVRQIAPGQTGKIA 419 , could be phosphorylated by phosphatidylinositol 3and/or 4-kinase (PIKKs). The Gln414 (Q) beside to the target Ser/Thr is critical for the substrate recognition. PIKKs are Ser/Thr atypical kinases that are found only in eukaryotes About 905 RFNGIG 910 , its C-terminal amino acids overlap with amino acids sequence 909 IGVTQNVL 916 that contains two motifs (phosphothreonine motif (TQ) and (T-x-x-I)) which Thr912 could be phosphorylated by PIKK family members, as described above for 408 RQIAPG 413 motif, and CaM-II kinase, respectively. As described above Thr415 is present in two patterns. When The RGD motif is the minimal peptide sequence used by many human viruses to SARS-CoV-2 is associated to significant cases of olfactory, taste, and/or chemesthesis disorders (OTCD) (von Bartheld et al., 2020). Because SARS-CoV-2 S protein contains two ARBMs (Fig. 1A , B) and some TRPs (TRPA1, TRPC1, TRPC3-7, and TRPV1-6) cation channels, implicated in OTC, have TRPs-ARDs, I hypothesized that the potential interaction of S-ARBMs and TRPs-ARDs could be responsible of OTCD. Some TRPs-ARDs are expressed in the olfactory epithelium, taste buds, trigeminal neurons in the oronasal cavity and vagal neurons in the trachea/lungs (Table 2) . Furthermore, this hypothesis is supported by works that have shown that (i) respiratory viruses interact with In order to interact with ARDs, 408 RQIAPG 413 and 905 RFNGIG 910 must be exposed at the surface of S protein. Indeed, these motifs are exposed in the surface of RBD and HR1 (PDB id: 3twr). Indeed, Frodock and Autodock Vina were able to produce a similar docking pose for each control protein with respect to its biological conformation in the cocrystallised protein-protein complex. Figure 3A shows that important amino acid residues R408 and G413 of 408 RQIAPG 413 in RBD interact with ankyrin repeat 6 (AR 6) of TRPA1. . Moreover, it had already been shown that the TRPV4 ARD interacts with the inositol head group of PI(4,5)P2, which negatively regulates the TRPV4 channel activity (Takahashi et al., 2014) . PI(4,5)P2 was located beside the AR 4 and interacts with the residues from AR 3-5 by its phosphate groups. The same result like RBD-TRPA1, shown above, was obtained with the docking of HR1 into ARD of TRPA1 (Fig. 3B) . To confirm the results of TRPA1-RBD and TRPA1-HR1 complexes, 408 RQIAPG 413 and 905 RFNGIG 910 peptides were used in blind docking. The docked peptides were localized in the same region where were localized RBD and HR1 in ARD of TRPA1 (Fig. 3C ). Structural alignment of docked 408 RQIAPG 413 and 905 RFNGIG 910 peptides with crystallised Tankyrase-2 AR-human SH3BP2 peptide (PDB id: 3twr) showed that these complexes were superposed (Fig. 3C) . This suggested the reliability of the blind docking. Of note, positively charged Arg408 and Arg905 of S-ARBMs interact with negatively charged region of electrostatic potential surface representation of the ARD region of TRPA1 (Fig. 3D ). Docking results of TRPV1 ARD with RBD and HR1 are represented in Figure 4 . The latter shows that RBD and HR1 interact with AR 3-4 of TRPV1 near cysteine 258 supposed to be implicated in the formation of inter-subunits disulfide bond (Tanaka et al., 2020) . Contrary to TRPA1 and 408 RQIAPG 413 and 905 RFNGIG 910 complexes that are superposed with the same orientation (N-to C-terminal), TRPV1-peptides complexes have opposed orientation (Fig. 4D ). The results of blind docking of S-ARBMs, 408 In the aim to find small molecules containing substituents that topologically and Designed ankyrin repeat proteins (DARPins) are small engineered nonimmunoglobulin AR proteins (14-21 kDa) (Stumpp et al., 2008) , usually composed of four to six AR motifs. They are considered as potential alternatives to monoclonal antibodies (mAbs) (Caputi and Navarra, 2020). DARPins are stable molecules with great affinity (picomolar), specificity and tissue penetration (Plückthun, 2015) . In addition, they can be administrated by different routes (oral, nasal, inhaled and topical). This is optimal to directly deliver of high dose of DARPins into oronasal cavity (where SARS-CoV-2 infection starts) and respiratory airways for both prophylactic and therapeutic protection in the early steps of the infection. For example, it has been reported that the entry of human immunodeficiency virus (HIV) into the host cell is blocked by DARPins. The latter compete with the HIV protein for the CD4 binding site on lymphocytes (Tomlinson et al., 2004 ). To defend the respiratory airways, TRPs induce airway constriction, sneezing and and TRPV4 antagonists could inhibit or mitigate inflammation. It has been suggested that other proteins such as neuropilin-1, PIKfyve kinase, Interestingly, the RGD motif is close to 408 RQIAPG 413 in a hot disordered loop ( 404 RGDEVRQIAPGQTGKIA 419 ), suggesting that it may participate in the internalization of S1 subunit. Different TRP channels are linked to viral infection (Table 3) It has been reported that Caucasians had a 3-6 times higher prevalence of OTCD The sensory neurons (nociceptors) and immune system work together to defend the To search probable short linear motifs (SLiMs), SARS-CoV-2 spike protein sequence was scanned with the eukaryotic linear motif (ELM) resource In the aim to find small molecules containing substituents that topologically and were refined by using FlexPepDock (London et al., 2011) , which allows full flexibility to the peptide and side-chain flexibility to the receptor. The electrostatic potential surface of hTRPA1 and hTRPV1 AR region was realized with PyMOL software (http://pymol.org/). I would like to thank the IBIS bioinformatics group for their help. The author declares that he has no conflicts of interest. 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