Publication:
Isolation and characterization of protein complexes of overexpressed human serum amyloid a from cytokines-induced HepG2 cells

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Authors
García Cortés, Carlos G.
Embargoed Until
Advisor
Parés Matos, Elsie I.
College
College of Arts and Sciences - Sciences
Department
Department of Chemistry
Degree Level
M.S.
Publisher
Date
2023-05-10
Abstract
Background and Objective: Serum Amyloid A (SAA) is an apolipoprotein involved in various physiological processes, including acute-phase response, cholesterol metabolism, retinol transport, and tumor pathogenesis. However, its exact biochemical role in many diseases has yet to be fully understood. Therefore, this research aims to identify the protein-protein interactions of SAA in human cells and to predict its biochemical role based on newly discovered complexes. Methods: HepG2 cells were induced with IL-1β, IL-6, Lipopolysaccharide, and Retinol, and the expression levels of SAA were measured using an ELISA assay. Protein complexes associated with SAA were isolated using the Co-Immunoprecipitation technique, and LC/MS/MS was used for characterization. Results: SAA was successfully overexpressed, and ten proteins were characterized by LC/MS/MS that may form complexes with SAA. Based on scientific publications, three of these proteins are associated with SAA. Discussion and Conclusions: The results of this research support some already established roles of SAA and provide new information related to its protein-protein interactions that could be used to develop new therapeutic interventions and diagnostic tools for diseases associated with SAA.

Antecedentes y objetivo: El amiloide sérico A (SAA) es una apolipoproteína involucrada en varios procesos fisiológicos, incluida la respuesta de fase aguda, el metabolismo del colesterol, el transporte de retinol y la patogénesis tumoral. Sin embargo, su papel bioquímico exacto en muchas enfermedades aún no se ha entendido por completo. Por lo tanto, esta investigación tiene como objetivo identificar las interacciones proteína-proteína de SAA en células humanas y predecir su papel bioquímico en base a complejos recién descubiertos. Métodos: Las células HepG2 se indujeron con IL-1β, IL-6, lipopolisacárido y retinol. Los niveles de proteína total y SAA se midieron mediante utilizando QUBIT 3.0 y ELISA, respectivamente. Los complejos proteicos asociados con SAA se aislaron utilizando la técnica de Co-Inmunoprecipitación, y se utilizó LC/MS/MS para la caracterización. Resultados: SAA se sobre expresó con éxito y diez proteínas se caracterizaron por LC/MS/MS que pueden formar complejos con SAA. Según publicaciones científicas, tres de estas proteínas están asociadas con SAA. Discusión y conclusiones: Los resultados de esta investigación fortalecen algunas funciones ya establecidas de SAA y brinda nueva información relacionada a sus interacciones proteína-proteína que podría utilizarse para desarrollar nuevas intervenciones terapéuticas y herramientas de diagnóstico para enfermedades asociadas con SAA.
Keywords
SAA,
acute-phase response,
Co-Immunoprecipitation,
mass spectrometry
Usage Rights
Except where otherwise noted, this item’s license is described as Attribution-NonCommercial-NoDerivatives 4.0 Internationl
Cite
García Cortés, C. G. (2023). Isolation and characterization of protein complexes of overexpressed human serum amyloid a from cytokines-induced HepG2 cells [Thesis]. Retrieved from https://hdl.handle.net/20.500.11801/3487