Abstract
Detection and purification of novel bioactive peptides from biological sources is a scientific task that led to a substantial number of important discoveries. One major laborious approach used is the repetitive stepwise separation of the test sample into several fractions followed by the determination of their bioactivity, until purity allows for sequence identification. We tested whether functional peptidomics, a combination of biological read-outs with differential peptide display (DPD) is a suitable strategy to isolate bioactive peptides at lower workload and with improved success. Additionally, we evaluated the use of DPD to monitor the processing status of proinsulin by inhibition of the insulin processing pathway. The rat insulinoma cell line INS-1 stimulated either with 2 mmol/l or 10 mmol/l glucose was used as model to generate differential peptide displays. In parallel, the bioactivity of the supernatants from the INS-1 cells was measured by glucose uptake and lipolysis assays using the adipocyte cell line 3T3-L1. We were able to quickly and elegantly trace the known activity of insulin to increase glucose uptake and inhibit lipolysis. Following re-chromatography of selected fractions, relevant peptides were identified by DPD and bioassays: the rat insulin-1 precursor and two different insulin peptides. We demonstrated in a semi-quantitative fashion that inhibition of proinsulin processing leads to accumulation of the insulin precursor, and reduced secretion of insulin-1. Thus, we conclude that DPD is an attractive support technology in peptide purification strategies aiming to identify bioactive compounds, and is superior to ELISA in discriminating between the processing status of insulin and its precursor.
Keywords: Adipocytes, Bioactivity, Differential Peptide Display, Functional Peptidomics, Glucose uptake, INS-1, Lipolysis, Proinsulin processing, Reversed-phase HPLC
Combinatorial Chemistry & High Throughput Screening
Title: The Concept of Functional Peptidomics for the Discovery of Bioactive Peptides in Cell Culture Models
Volume: 8 Issue: 8
Author(s): Marco M. Jost, Petra Budde, Harald Tammen, Rudiger Hess, Markus Kellmann, Imke Schulte and Horst Rose
Affiliation:
Keywords: Adipocytes, Bioactivity, Differential Peptide Display, Functional Peptidomics, Glucose uptake, INS-1, Lipolysis, Proinsulin processing, Reversed-phase HPLC
Abstract: Detection and purification of novel bioactive peptides from biological sources is a scientific task that led to a substantial number of important discoveries. One major laborious approach used is the repetitive stepwise separation of the test sample into several fractions followed by the determination of their bioactivity, until purity allows for sequence identification. We tested whether functional peptidomics, a combination of biological read-outs with differential peptide display (DPD) is a suitable strategy to isolate bioactive peptides at lower workload and with improved success. Additionally, we evaluated the use of DPD to monitor the processing status of proinsulin by inhibition of the insulin processing pathway. The rat insulinoma cell line INS-1 stimulated either with 2 mmol/l or 10 mmol/l glucose was used as model to generate differential peptide displays. In parallel, the bioactivity of the supernatants from the INS-1 cells was measured by glucose uptake and lipolysis assays using the adipocyte cell line 3T3-L1. We were able to quickly and elegantly trace the known activity of insulin to increase glucose uptake and inhibit lipolysis. Following re-chromatography of selected fractions, relevant peptides were identified by DPD and bioassays: the rat insulin-1 precursor and two different insulin peptides. We demonstrated in a semi-quantitative fashion that inhibition of proinsulin processing leads to accumulation of the insulin precursor, and reduced secretion of insulin-1. Thus, we conclude that DPD is an attractive support technology in peptide purification strategies aiming to identify bioactive compounds, and is superior to ELISA in discriminating between the processing status of insulin and its precursor.
Export Options
About this article
Cite this article as:
Jost M. Marco, Budde Petra, Tammen Harald, Hess Rudiger, Kellmann Markus, Schulte Imke and Rose Horst, The Concept of Functional Peptidomics for the Discovery of Bioactive Peptides in Cell Culture Models, Combinatorial Chemistry & High Throughput Screening 2005; 8 (8) . https://dx.doi.org/10.2174/138620705774962445
DOI https://dx.doi.org/10.2174/138620705774962445 |
Print ISSN 1386-2073 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5402 |
Call for Papers in Thematic Issues
Artificial Intelligence Methods for Biomedical, Biochemical and Bioinformatics Problems
Recently, a large number of technologies based on artificial intelligence have been developed and applied to solve a diverse range of problems in the areas of biomedical, biochemical and bioinformatics problems. By utilizing powerful computing resources and massive amounts of data, methods based on artificial intelligence can significantly improve the ...read more
Eco-friendly Agents for Biological Control of Pathogenic Diseases
The discovery of an alternative biological approach to disease management includes work on medicinal products derived from natural sources as a starting point for the development of eco-friendly agents for these diseases and the injuries they cause, as well as reducing human contact with hazardous chemicals and their residues. We ...read more
Emerging trends in diseases mechanisms, noble drug targets and therapeutic strategies: focus on immunological and inflammatory disorders
Recently infectious and inflammatory diseases have been a key concern worldwide due to tremendous morbidity and mortality world Wide. Recent, nCOVID-9 pandemic is a good example for the emerging infectious disease outbreak. The world is facing many emerging and re-emerging diseases out breaks at present however, there is huge lack ...read more
Exploring Spectral Graph Theory in Combinatorial Chemistry
Scope of the Thematic Issue: Combinatorial chemistry involves the synthesis and analysis of a large number of diverse compounds simultaneously. Traditional methods rely on brute force experimentation, which can be time-consuming and resource-intensive. Spectral Graph Theory, a branch of mathematics dealing with the properties of graphs in relation to the ...read more
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Advances in Treating the Ischaemic Diabetic Foot
Current Vascular Pharmacology Preface
Current Pharmaceutical Design Multiple Drug Resistance Associated with Function of ABC-Transporters in Diabetes Mellitus: Molecular Mechanism and Clinical Relevance
Infectious Disorders - Drug Targets The PIK3CA Gene as a Mutated Target for Cancer Therapy
Current Cancer Drug Targets Protein Interaction Networks in Metallo Proteins and Docking Approaches of Metallic Compounds with TIMP and MMP in Control of MAPK Pathway
Letters in Drug Design & Discovery Role of Pancreatic β-Cell Death and Cell Death-Associated Inflammation in Diabetes
Current Molecular Medicine Update on the Evidence of Statin Treatment to Reduce Plaque Vulnerability
Drug Metabolism Letters A Review on Screening Models for Potential Therapeutic Candidates and Targets Against SARS-CoV-2
Current Drug Targets The Interactions of Anti-Cancer Drugs Approved in the Last Decade in the United States with Membrane Transporters
Anti-Cancer Agents in Medicinal Chemistry The Role of Insertion Allele of Angiotensin Converting Enzyme Gene in Higher Endurance Efficiency and Some Aspects of Pathophysiological and Drug Effects
Current Medicinal Chemistry Gender Differences in Autonomic Control of the Cardiovascular System
Current Pharmaceutical Design Safety and Tolerability of Agomelatine: Focus on Hepatotoxicity
Current Drug Metabolism Buxus sempervirens L. Improves Lipid Profile in Diabetic Rats
Cardiovascular & Hematological Disorders-Drug Targets Macrophage Migration Inhibitory Factor: A Therapeutic Target Across Inflammatory Diseases
Inflammation & Allergy - Drug Targets (Discontinued) Meet Our Editorial Board Member
Current Drug Targets A Novel SCN9A Mutation (F826Y) in Primary Erythromelalgia Alters the Excitability of Nav1.7
Current Molecular Medicine Endothelium as a Therapeutic Target in Diabetes Mellitus: From Basic Mechanisms to Clinical Practice
Current Medicinal Chemistry Computational & Statistical Methodologies to Identify Biomarkers in Cancer
Current Cancer Therapy Reviews Genistein Aglycone: A Dual Mode of Action Anti-Osteoporotic Soy Isoflavone Rebalancing Bone Turnover Towards Bone Formation
Current Medicinal Chemistry Targeting Transient Receptor Potential Canonical Channels for Diseases of the Nervous System
Current Drug Targets