Project 11

Project 11: Enrichment, identification and characterization of urinary proteases.

Principal Investigator: Professor Harry Holthofer

Project Description:

Chronic kidney disease (CKD) is a major cause of morbidity and mortality which places a significant financial burden on Ireland's healthcare system. A key consideration in combating the ever worsening disease epidemic is having a better understanding of the disease mechanisms. This includes identifying suitable biomarkers for permitting early detection and risk assessment, and identifying novel therapeutic routes through the development of an understanding of key target molecules and pathways. Urine is regarded as one of the most promising sources of easily obtainable bodily fluids which can be collected non-invasively and in large, thus providing an attractive alternative to blood/serum as a source of disease indicators. Despite the normally low urinary protein concentration, large-scale proteomic profiling has revealed over 2,300 different protein gene products.

The number of urinary protein identification studies has been increasing since the methodological approach aimed to discover proteins express in very low amount also known as "rare proteome". Among these, glycoproteins play a double role both with their protein backbone and their glycan moieties. Glycoproteins have a significant biomedical interest because modifications of glycosylation domains have recently been established as early markers in the development of diseases. This project will focus on two main areas, namely (1) isolation of glycoproteins from the urine and (2) their biochemical characterization.

Benefits for the Student:

During this time the student will:

Gain support from a well-established research group with expertise in proteomics, glycomics and in methods of general in protein chemistry. Furthermore, the student will develop key skills in good laboratory practice, and an understanding of the clinical relevance of CKD and how this global epidemic is currently being tackled.

Develop expertise in traditional and high-throughput methodologies for the isolation of urinary proteases and protease inhibitors.

Be introduced to proteomic and glycomic techniques that allow analysis of proteins and carbohydrates, respectively.

Be encouraged to participate in group meetings and research update seminars, thus improving presentation and troubleshooting skills.
Be included as a collaborator on subsequent manuscripts resulting from their contribution in the laboratory.
How the project fits in with the overall objective of the research group:

Considerable progress has been made in the analysis of urinary proteome especially at the proteomic level but sub proteome expression at low levels remains to be characterized. This work is ongoing in our laboratory. Furthermore, the analysis of urinary proteases and their activity is of considerable clinical importance in providing another, potentially hugely important aspect of disease biology and, hence, designing therapeutic strategies in collaboration with our European partners.

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	Project 11: Enrichment, identification and characterization of urinary proteases. Principal Investigator: Professor Harry Holthofer Project Description: Chronic kidney disease (CKD) is a major cause of morbidity and mortality which places a significant financial burden on Ireland's healthcare system. A key consideration in combating the ever worsening disease epidemic is having a better understanding of the disease mechanisms. This includes identifying suitable biomarkers for permitting early detection and risk assessment, and identifying novel therapeutic routes through the development of an understanding of key target molecules and pathways. Urine is regarded as one of the most promising sources of easily obtainable bodily fluids which can be collected non-invasively and in large, thus providing an attractive alternative to blood/serum as a source of disease indicators. Despite the normally low urinary protein concentration, large-scale proteomic profiling has revealed over 2,300 different protein gene products. The number of urinary protein identification studies has been increasing since the methodological approach aimed to discover proteins express in very low amount also known as "rare proteome". Among these, glycoproteins play a double role both with their protein backbone and their glycan moieties. Glycoproteins have a significant biomedical interest because modifications of glycosylation domains have recently been established as early markers in the development of diseases. This project will focus on two main areas, namely (1) isolation of glycoproteins from the urine and (2) their biochemical characterization. Benefits for the Student: During this time the student will: Gain support from a well-established research group with expertise in proteomics, glycomics and in methods of general in protein chemistry. Furthermore, the student will develop key skills in good laboratory practice, and an understanding of the clinical relevance of CKD and how this global epidemic is currently being tackled. Develop expertise in traditional and high-throughput methodologies for the isolation of urinary proteases and protease inhibitors. Be introduced to proteomic and glycomic techniques that allow analysis of proteins and carbohydrates, respectively. Be encouraged to participate in group meetings and research update seminars, thus improving presentation and troubleshooting skills. Be included as a collaborator on subsequent manuscripts resulting from their contribution in the laboratory. How the project fits in with the overall objective of the research group: Considerable progress has been made in the analysis of urinary proteome especially at the proteomic level but sub proteome expression at low levels remains to be characterized. This work is ongoing in our laboratory. Furthermore, the analysis of urinary proteases and their activity is of considerable clinical importance in providing another, potentially hugely important aspect of disease biology and, hence, designing therapeutic strategies in collaboration with our European partners. Back