Purification and distribution of HSP90 in sheep liver tissue
Journal name: World Journal of Pharmaceutical Research
Original article title: Subcellular distribution and purification of heat shock protein 90 in sheep liver tissue
The WJPR includes peer-reviewed publications such as scientific research papers, reports, review articles, company news, thesis reports and case studies in areas of Biology, Pharmaceutical industries and Chemical technology while incorporating ancient fields of knowledge such combining Ayurveda with scientific data.
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Summary of article contents:
Introduction
Heat shock proteins (HSPs) are highly conserved proteins that play vital roles in cellular functions, especially under stress conditions. The 90 kDa heat shock protein (HSP90) is one of the most abundant proteins found in cells, functioning primarily as a molecular chaperone. This study, conducted by Rifat Ara and colleagues, focuses on the purification and subcellular distribution of HSP90 in sheep liver tissue. It employs various biochemical techniques, including DEAE-cellulose chromatography and immunoblotting, to analyze the presence and behavior of HSP90 across different cellular compartments.
Role of Heat Shock Proteins
Heat shock proteins are crucial for cellular homeostasis, helping in the proper folding and stabilization of other proteins. Particularly, HSP90 is associated with a variety of intracellular proteins, including kinases and steroid receptors, which are essential for cellular signaling and function. The study notes that HSP90 not only activates and stabilizes target proteins in inactive states but also assists in their translocation across cellular membranes. By understanding HSP90’s molecular mechanisms, researchers can gain insights into its significance in cellular stress response and overall protein management under both normal and stress conditions.
Methodology for Protein Purification
The researchers implemented a detailed methodology for isolating and purifying HSP90 from sheep liver. Initial steps involved homogenizing liver tissue and subjecting it to differential centrifugation to obtain cytosolic proteins. Subsequent purification utilized ammonium sulfate precipitation followed by anion-exchange chromatography. The effectiveness of the purification process was confirmed through SDS-PAGE, revealing a predominant band corresponding to HSP90. The approach emphasizes the use of mammalian tissues over cultured cells for obtaining significant yields of protein, which is critical for further functional studies.
Subcellular Localization of HSP90
Using immunoblotting techniques, the study further examined the subcellular distribution of HSP90 within different fractions of liver tissue. The results demonstrated that HSP90 is widely distributed across various cellular fractions—cytosolic, nuclear, and mitochondrial—highlighting its multifunctional capacities as a chaperone. Immunoblotting revealed a consistent presence of HSP90 at an apparent molecular weight of 90 kDa, affirming its role as a key participant in numerous biological processes across cellular compartments.
Conclusion
In conclusion, this study successfully demonstrated the isolation and characterization of HSP90 from sheep liver tissue. The findings contribute valuable insights into the functional dynamics and significance of HSP90, particularly its role as a crucial molecular chaperone in maintaining protein integrity and stability across various subcellular environments. As the research advances, it is expected that further exploration into HSP90's functions will enhance understanding of its implications in stress responses and cellular health in mammals.
Original source:
This page is merely a summary which is automatically generated hence you should visit the source to read the original article which includes the author, publication date, notes and references.
Rifat Ara, Ruqaya Aziz, Shazia Nazir, Aeijaz Ul Noor, Khurshid I Andrabi
World Journal of Pharmaceutical Research:
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Full text available for: Subcellular distribution and purification of heat shock protein 90 in sheep liver tissue
Source type: An International Peer Reviewed Journal for Pharmaceutical and Medical and Scientific Research
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FAQ section (important questions/answers):
What is Heat Shock Protein 90 (HSP90) and its function?
HSP90 is a highly conserved molecular chaperone that assists in protein maturation, including folding and unfolding, and stabilizes target proteins in an inactive state, crucial for normal cellular function.
How was HSP90 purified from sheep liver tissue?
HSP90 was purified using DEAE-cellulose chromatography and ammonium sulfate precipitation techniques, yielding a significant amount of the protein for further studies.
What methods were used to analyze the presence of HSP90?
The presence of HSP90 was analyzed using Western blotting after protein extraction and separation by SDS-PAGE, confirming its molecular weight and distribution in various cellular fractions.
What were the sources of liver tissue used in the study?
Liver tissue was sourced from healthy sheep obtained immediately after slaughter from a local abattoir to ensure freshness for the experiments.
What significance does HSP90 have in liver tissue?
HSP90 exhibits a ubiquitous distribution across cellular fractions, indicating its essential role in various cellular processes and its importance in liver tissue function.
What role do heat shock proteins have under stress conditions?
Under stress conditions, heat shock proteins like HSP90 are induced to protect cells by assisting in the repair and proper folding of denatured proteins, thereby promoting cell survival.
Glossary definitions and references:
Scientific and Ayurvedic Glossary list for “Purification and distribution of HSP90 in sheep liver tissue”. This list explains important keywords that occur in this article and links it to the glossary for a better understanding of that concept in the context of Ayurveda and other topics.
1) Kashmir:
Kashmir is a region in northern India, significant in this study as the location of SKIMS Medical College and the University of Kashmir, where the research was conducted. The context emphasizes the geographical and institutional background relevant to the subcellular distribution and purification of heat shock protein 90 in sheep liver tissue.
2) Purification:
Purification refers to the process of isolating a specific protein, HSP90 in this study, from other cellular components. The research highlights various chromatographic techniques employed to achieve a high yield and purity of HSP90 from sheep liver, essential for studying its functional properties and scientific analysis.
3) India:
India is the country where the research was conducted, specifically in the Jammu and Kashmir region. The mention of India underlines the geographical significance and the scientific community involved in understanding the molecular biology of heat shock proteins in livestock, contributing to the advancement of biotechnology.
4) Srinagar:
Srinagar is the summer capital of Jammu and Kashmir and is central to the research setting as the location of the departments involved in the study. It highlights the local institutional support and resources available for conducting biomedical research, specifically focusing on the examination of heat shock proteins.
5) Powder:
Powder, in this context, refers to carnation milk powder used as a blocking agent in the Western blotting technique outlined in the methods. Its role is vital for reducing non-specific binding, thereby enhancing the specificity and clarity of the protein detection associated with HSP90.
6) Study (Studying):
The study is a systematic investigation aimed at exploring the subcellular distribution and purification of heat shock protein 90 in sheep liver. It contributes to the body of knowledge regarding the role of HSPs in cellular function and has implications for understanding stress responses in various organisms.
7) Milk:
Milk is mentioned in the context of using milk powder for blocking during the Western blotting procedure. The use of milk in biochemical assays is a common practice to prevent non-specific binding of antibodies, indicating attention to methodological details in protein analysis.
8) Transformation (Transform, Transforming):
Transforming relates to the protein-protein interactions that heat shock proteins like HSP90 facilitate, specifically in the context of cellular processes. The study's discussion of transforming proteins highlights the functional roles of HSP90 in modulating the activity and localization of various target proteins within cells.
9) Discussion:
Discussion refers to the analytical dialogue presented in the study regarding the significance of the results on the distribution and functions of HSP90. It contextualizes the findings within the broader scientific framework, contributing to the understanding of chaperone proteins in cellular biology.
10) Similarity:
Similarity pertains to the comparative analysis of protein sequences and functions, particularly how HSP90 demonstrates conservation across species. The study emphasizes the evolutionary importance of HSP90, reflecting its fundamental role in cellular stress responses, thus carrying implications for evolutionary biology studies.
11) Evolution:
Evolution underlines the concept that heat shock proteins are conserved across different species due to their fundamental role in cellular processes. The study emphasizes the evolutionary significance of HSP90, revealing how vital such proteins are for maintaining homeostasis during environmental stressors across species.
12) Medicine:
Medicine is central to the implications of the research, as understanding heat shock proteins like HSP90 can lead to insights into disease mechanisms and therapeutic strategies. The findings have the potential to influence medical research aimed at stress-related diseases, cancer, and other health conditions.
13) Disease:
Disease references the broader health implications associated with heat shock proteins, particularly their role in protein misfolding and cellular stress responses. The research contributes to a better understanding of how HSP90 may be involved in various disease processes, pointing towards future therapeutic targets.
14) Science (Scientific):
Science embodies the systematic study and application of knowledge in the field of molecular biology explored in this research. The investigation into heat shock proteins represents the intersection of scientific inquiry, experimentation, and the pursuit of understanding cellular and molecular mechanisms in living organisms.
15) Nature:
Nature references the biological and physical phenomena observed in the natural world, which underpin the study of heat shock proteins. Understanding HSP90's roles in biological systems reflects a fundamental aspect of scientific inquiry into nature's mechanisms, influencing developments in biotechnology and environmental biology.
16) Avila:
Avila is possibly mentioned in reference to previous studies outlining the interactions and roles that proteins such as HSP70 and HSP90 play in cellular functions. Such references anchor the study within the broader context of existing literature and methods used for studying heat shock proteins.
17) Beta:
Beta refers to beta-mercaptoethanol, a reducing agent used in the study's protein assays to maintain reduced conditions during protein extraction and analysis. Its role is crucial in preventing the formation of disulfide bonds, thus ensuring accurate protein characterization and detection in Western blotting.
18) Salt (Salty):
Salt, particularly in the context of ammonium sulfate precipitation and anion-exchange chromatography, is an essential component for protein purification. It is used to precipitate proteins and maintain ionic strength during chromatography, critical for isolating HSP90 effectively from liver tissue extracts.
19) Post:
Post refers to the 'post-nuclear supernatant', indicating the fraction of tissue homogenate remaining after centrifugation. This term underscores the methodology of separating cellular components to isolate specific fractions for further analysis of HSP90 distribution and presence in different cellular organelles.
20) Viru:
Viru does not appear explicitly in the text provided. If it refers to a concept or is a misinterpretation of another term, further context is needed to provide a relevant explanation. Thus, no definition can be appropriately generated without additional information.