دورية أكاديمية
Identification of transient receptor potential melastatin 3 proteotypic peptides employing an efficient membrane protein extraction method for natural killer cells.
| العنوان: | Identification of transient receptor potential melastatin 3 proteotypic peptides employing an efficient membrane protein extraction method for natural killer cells. |
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| المؤلفون: | Magawa CT; National Centre for Neuroimmunology and Emerging Diseases, Menzies Health Institute Queensland, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia.; Consortium Health International for Myalgic Encephalomyelitis, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia., Eaton-Fitch N; National Centre for Neuroimmunology and Emerging Diseases, Menzies Health Institute Queensland, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia.; Consortium Health International for Myalgic Encephalomyelitis, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia., Balinas C; National Centre for Neuroimmunology and Emerging Diseases, Menzies Health Institute Queensland, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia.; Consortium Health International for Myalgic Encephalomyelitis, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia., Sasso EM; National Centre for Neuroimmunology and Emerging Diseases, Menzies Health Institute Queensland, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia.; Consortium Health International for Myalgic Encephalomyelitis, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia.; School of Pharmacy and Medical Sciences, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia., Thapaliya K; National Centre for Neuroimmunology and Emerging Diseases, Menzies Health Institute Queensland, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia.; Consortium Health International for Myalgic Encephalomyelitis, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia., Barnden L; National Centre for Neuroimmunology and Emerging Diseases, Menzies Health Institute Queensland, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia.; Consortium Health International for Myalgic Encephalomyelitis, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia., Maksoud R; National Centre for Neuroimmunology and Emerging Diseases, Menzies Health Institute Queensland, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia.; Consortium Health International for Myalgic Encephalomyelitis, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia.; School of Pharmacy and Medical Sciences, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia., Weigel B; National Centre for Neuroimmunology and Emerging Diseases, Menzies Health Institute Queensland, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia.; Consortium Health International for Myalgic Encephalomyelitis, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia.; School of Pharmacy and Medical Sciences, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia., Rudd PA; Institute for Glycomics, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia., Herrero LJ; Institute for Glycomics, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia., Marshall-Gradisnik S; National Centre for Neuroimmunology and Emerging Diseases, Menzies Health Institute Queensland, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia.; Consortium Health International for Myalgic Encephalomyelitis, Griffith University, Gold Coast Campus, Gold Coast, Qld, Australia. |
| المصدر: | Frontiers in physiology [Front Physiol] 2022 Sep 23; Vol. 13, pp. 947723. Date of Electronic Publication: 2022 Sep 23 (Print Publication: 2022). |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Frontiers Research Foundation Country of Publication: Switzerland NLM ID: 101549006 Publication Model: eCollection Cited Medium: Print ISSN: 1664-042X (Print) Linking ISSN: 1664042X NLM ISO Abbreviation: Front Physiol Subsets: PubMed not MEDLINE |
| أسماء مطبوعة: | Original Publication: Lausanne : Frontiers Research Foundation |
| مستخلص: | Introduction: Mutations and misfolding of membrane proteins are associated with various disorders, hence they make suitable targets in proteomic studies. However, extraction of membrane proteins is challenging due to their low abundance, stability, and susceptibility to protease degradation. Given the limitations in existing protocols for membrane protein extraction, the aim of this investigation was to develop a protocol for a high yield of membrane proteins for isolated Natural Killer (NK) cells. This will facilitate genetic analysis of membrane proteins known as transient receptor potential melastatin 3 (TRPM3) ion channels in myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) research. Methods: Two protocols, internally identified as Protocol 1 and 2, were adapted and optimized for high yield protein extraction. Protocol 1 utilized ultrasonic and salt precipitation, while Protocol 2 implemented a detergent and chloroform/methanol approach. Protein concentrations were determined by the Pierce Bicinchoninic Acid (BCA) and the Bio-Rad DC (detergent compatible) protein assays according to manufacturer's recommendation. Using Protocol 2, protein samples were extracted from NK cells of n = 6 healthy controls (HC) and n = 4 ME/CFS patients. In silico tryptic digest and enhanced signature peptide (ESP) predictor were used to predict high-responding TRPM3 tryptic peptides. Trypsin in-gel digestion was performed on protein samples loaded on SDS-PAGE gels (excised at 150-200 kDa). A liquid chromatography-multiple reaction monitoring (LC-MRM) method was optimized and used to evaluate the detectability of TRPM3 n = 5 proteotypic peptides in extracted protein samples. Results: The detergent-based protocol protein yield was significantly higher ( p < 0.05) compared with the ultrasonic-based protocol. The Pierce BCA protein assay showed more reproducibility and compatibility compared to the Bio-Rad DC protein assay. Two high-responding tryptic peptides (GANASAPDQLSLALAWNR and QAILFPNEEPSWK) for TRPM3 were detectable in n = 10 extracted protein samples from NK cells isolated from HC and ME/CFS patients. Conclusion: A method was optimized for high yield protein extraction from human NK cells and for the first time TRPM3 proteotypic peptides were detected using LC-MRM. This new method provides for future research to assess membrane protein structural and functional relationships, particularly to facilitate proteomic investigation of TRPM3 ion channel isoforms in NK cells in both health and disease states, such as ME/CFS. Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2022 Magawa, Eaton-Fitch, Balinas, Sasso, Thapaliya, Barnden, Maksoud, Weigel, Rudd, Herrero and Marshall-Gradisnik.) |
| References: | J Biol Chem. 2012 Oct 26;287(44):36663-72. (PMID: 22961981) J Clin Invest. 2020 Mar 2;130(3):1491-1505. (PMID: 31830003) Methods Enzymol. 2009;463:619-29. (PMID: 19892195) Pflugers Arch. 2010 Jul;460(2):437-50. (PMID: 20127491) Asian Pac J Allergy Immunol. 2019 Jun;37(2):102-108. (PMID: 29981562) Biology (Basel). 2021 Mar 22;10(3):. (PMID: 33809831) Hum Genomics. 2020 Feb 18;14(1):7. (PMID: 32070426) Pharmaceutics. 2019 Jan 18;11(1):. (PMID: 30669330) Sci STKE. 2005 Aug 02;2005(295):re8. (PMID: 16077087) Biology (Basel). 2020 Nov 16;9(11):. (PMID: 33207666) Biochem Soc Trans. 2019 Feb 28;47(1):47-61. (PMID: 30559269) Biol Res. 2016 May 31;49(1):27. (PMID: 27245705) J Eval Clin Pract. 2012 Feb;18(1):25-31. (PMID: 21029269) Eur J Hum Genet. 2019 Oct;27(10):1611-1618. (PMID: 31278393) J Clin Invest. 2004 Dec;114(11):1612-23. (PMID: 15578093) Methods Enzymol. 2009;463:73-95. (PMID: 19892168) Clin Exp Immunol. 2017 Feb;187(2):284-293. (PMID: 27727448) J Biol Chem. 2003 Jun 13;278(24):21493-501. (PMID: 12672799) Mol Cell Proteomics. 2013 May;12(5):1099-114. (PMID: 23315794) Front Cell Dev Biol. 2021 Feb 26;9:635659. (PMID: 33732703) Annu Rev Physiol. 2006;68:619-47. (PMID: 16460286) Front Immunol. 2019 Oct 31;10:2545. (PMID: 31736966) Methods Mol Biol. 2016;1362:63-76. (PMID: 26519169) Mol Syst Biol. 2008;4:222. (PMID: 18854821) Front Immunol. 2021 Jul 13;12:687806. (PMID: 34326841) Methods Mol Biol. 2012;869:29-35. (PMID: 22585474) J Biol Chem. 2005 Jun 10;280(23):22540-8. (PMID: 15824111) Nat Biotechnol. 2009 Feb;27(2):190-8. (PMID: 19169245) EMBO Rep. 2008 Jun;9(6):512-5. (PMID: 18451877) J Intern Med. 2011 Oct;270(4):327-38. (PMID: 21777306) J Med Chem. 2013 Feb 14;56(3):593-624. (PMID: 23121096) Anal Bioanal Chem. 2022 Feb;414(5):1933-1947. (PMID: 34997251) Electrophoresis. 2013 Mar;34(6):809-17. (PMID: 23334993) Materials (Basel). 2016 May 18;9(5):. (PMID: 28773508) Cell Calcium. 2008 Nov;44(5):492-506. (PMID: 18436303) J Transl Med. 2021 Feb 17;19(1):81. (PMID: 33596913) J Biol Chem. 2003 Jun 6;278(23):20890-7. (PMID: 12672827) Methods Enzymol. 2014;541:117-40. (PMID: 24674067) Nat Methods. 2012 May 30;9(6):555-66. (PMID: 22669653) Endocrinol Metab (Seoul). 2017 Mar;32(1):18-22. (PMID: 28256117) Annu Rev Biochem. 2007;76:387-417. (PMID: 17579562) Nat Rev Nephrol. 2009 Aug;5(8):441-9. (PMID: 19546862) Methods Enzymol. 2009;463:285-303. (PMID: 19892178) Mol Med. 2018 Aug 14;24(1):44. (PMID: 30134818) Cytotechnology. 2009 Dec;61(3):153-9. (PMID: 20072854) Cell Calcium. 2005 Sep-Oct;38(3-4):233-52. (PMID: 16098585) |
| فهرسة مساهمة: | Keywords: TRP channels; TRPM3; TRPM3 proteotypic peptides; calcium signaling; membrane protein extraction; natural killer cells |
| تواريخ الأحداث: | Date Created: 20221010 Latest Revision: 20221011 |
| رمز التحديث: | 20240628 |
| مُعرف محوري في PubMed: | PMC9540229 |
| DOI: | 10.3389/fphys.2022.947723 |
| PMID: | 36213251 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1664-042X |
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| DOI: | 10.3389/fphys.2022.947723 |