Combining complementary multiplex extraction chemistries enhances proteome coverage and analytical insights in tiny insects: A study on sweet potato whitefly Bemisia tabaci.
Saved in:
| Title: | Combining complementary multiplex extraction chemistries enhances proteome coverage and analytical insights in tiny insects: A study on sweet potato whitefly Bemisia tabaci. |
|---|---|
| Authors: | Kolanchi P; Department of Agricultural Entomology, Centre for Plant Protection Studies, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India., Sanivarapu H; Cell & Molecular Biology and Trait Engineering, International Crops Research Institute for the Semi-Arid Tropics, Hyderabad, India., Marimuthu M; Department of Agricultural Entomology, Centre for Plant Protection Studies, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India. Electronic address: muruganmarimuthu@tnau.ac.in., Kalenahalli Y; Cell & Molecular Biology and Trait Engineering, International Crops Research Institute for the Semi-Arid Tropics, Hyderabad, India., Onkarappa D; Department of Agricultural Entomology, Centre for Plant Protection Studies, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India., Jagdish J; Crop Protection & Seed Health cluster, International Crops Research Institute for the Semi-Arid Tropics, Hyderabad, India., Venkatasamy B; Department of Agricultural Entomology, Centre for Plant Protection Studies, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India., Muthu A; Department of Agricultural Entomology, Centre for Plant Protection Studies, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India. |
| Source: | International journal of biological macromolecules [Int J Biol Macromol] 2025 Dec; Vol. 333 (Pt 1), pp. 148741. Date of Electronic Publication: 2025 Nov 07. |
| Publication Type: | Journal Article |
| Language: | English |
| Journal Info: | Publisher: Elsevier Country of Publication: Netherlands NLM ID: 7909578 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1879-0003 (Electronic) Linking ISSN: 01418130 NLM ISO Abbreviation: Int J Biol Macromol Subsets: MEDLINE |
| Imprint Name(s): | Publication: Amsterdam : Elsevier Original Publication: Guildford, Eng., IPC Science and Technology Press. |
| MeSH Terms: | Hemiptera*/metabolism , Hemiptera*/chemistry , Proteome*/isolation & purification , Proteomics*/methods , Insect Proteins*/isolation & purification , Insect Proteins*/metabolism , Ipomoea batatas*/parasitology, Animals ; Tandem Mass Spectrometry ; Chromatography, Liquid |
| Abstract: | Advancements in protein extraction methodologies are essential for enhancing insect proteomics, especially in sap-feeding small insects, where high lipid content and limited tissue volume impede efficient protein recovery. This study systematically assessed seven extraction chemistries, lysis buffer (LB), dissolution buffer (DB), TRIzol®, trifluoroacetic acid (TFA), trichloroacetic acid (TCA), TCA-phenol, and phenol- under varying pH conditions to determine the most effective workflow for comprehensive proteomic profiling of Bemisia tabaci. Qualitative two-dimensional electrophoresis (2-DE) analysis indicated that phenol and TCA-phenol methods maintained protein integrity across a wide molecular weight range, while acidic conditions enhanced the resolution of low-abundance proteins. Quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses demonstrated that the TCA-phenol method yielded the highest protein recovery and reproducibility (coefficient of variation = 12.6 %), identifying 1010 proteins with balanced representation of soluble and membrane fractions. Principal component and overlap analyses confirmed distinct, method-specific proteomic signatures, with TCA-phenol and TFA achieving the most extensive proteome coverage. Functional annotation revealed that DB and LB enriched primary metabolic and biosynthetic processes, whereas the TCA-phenol method provided the broadest functional spectrum, capturing detoxification, energy metabolism, and stress-related proteins. Overall, this research illustrates that no single extraction chemistry is universally optimal; rather, combining complementary methods enhances proteome coverage. These findings establish a standardized comparative framework for small-insect proteomics and offer valuable methodological insights to advance research in insect physiology, stress adaptation, and ecological interactions. (Copyright © 2025 Elsevier B.V. All rights reserved.) |
| Competing Interests: | Declaration of competing interest The authors declare that they have no competing interests. |
| Contributed Indexing: | Keywords: 2-DE; Bemisia tabaci; Benefit-to-cost ratio; LC–MS/MS; SDS-PAGE; TCA-phenol |
| Substance Nomenclature: | 0 (Proteome) 0 (Insect Proteins) |
| Entry Date(s): | Date Created: 20251109 Date Completed: 20251202 Latest Revision: 20251202 |
| Update Code: | 20251202 |
| DOI: | 10.1016/j.ijbiomac.2025.148741 |
| PMID: | 41207581 |
| Database: | MEDLINE |
Full Text Finder 
Nájsť tento článok vo Web of Science | Abstract: | Advancements in protein extraction methodologies are essential for enhancing insect proteomics, especially in sap-feeding small insects, where high lipid content and limited tissue volume impede efficient protein recovery. This study systematically assessed seven extraction chemistries, lysis buffer (LB), dissolution buffer (DB), TRIzol®, trifluoroacetic acid (TFA), trichloroacetic acid (TCA), TCA-phenol, and phenol- under varying pH conditions to determine the most effective workflow for comprehensive proteomic profiling of Bemisia tabaci. Qualitative two-dimensional electrophoresis (2-DE) analysis indicated that phenol and TCA-phenol methods maintained protein integrity across a wide molecular weight range, while acidic conditions enhanced the resolution of low-abundance proteins. Quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses demonstrated that the TCA-phenol method yielded the highest protein recovery and reproducibility (coefficient of variation = 12.6 %), identifying 1010 proteins with balanced representation of soluble and membrane fractions. Principal component and overlap analyses confirmed distinct, method-specific proteomic signatures, with TCA-phenol and TFA achieving the most extensive proteome coverage. Functional annotation revealed that DB and LB enriched primary metabolic and biosynthetic processes, whereas the TCA-phenol method provided the broadest functional spectrum, capturing detoxification, energy metabolism, and stress-related proteins. Overall, this research illustrates that no single extraction chemistry is universally optimal; rather, combining complementary methods enhances proteome coverage. These findings establish a standardized comparative framework for small-insect proteomics and offer valuable methodological insights to advance research in insect physiology, stress adaptation, and ecological interactions.<br /> (Copyright © 2025 Elsevier B.V. All rights reserved.) |
|---|---|
| ISSN: | 1879-0003 |
| DOI: | 10.1016/j.ijbiomac.2025.148741 |