Development of iCrystal System for Rapid Crystallization of Biological Macromolecules

DESCRIPTION provided by applicant Biological macromolecules are the machinery of life and understanding their function helps scientists to develop new drug treatments that target specific human diseases In this regard crystallization is routinely employed for the understanding of the molecular structures and the interactions of proteins with other biological and non biological materials Despite the existence of a plethora of crystallization techniques there is still a need for a technique that affords for better control over the crystallization procss in terms of producing high quality crystals of peptides and proteins in a significantly shorter tim scale In this STTR Phase I proposal we will construct a prototype crystallization instrument based on the metal assisted and microwave assisted evaporative crystallization MA MAEC technique for the rapid crystallization of peptides and proteins in minutes or hours total crystallization time which typically can take up to several weeks to complete using conventional crystallization techniques In this regard we have chosen Amyloid precursor protein APP and its components as biological macromolecules of interest APP is most commonly studied as the precursor molecule whose proteolysis generates beta amyloid A peptide whose amyloid fibrillar form is the primary component of amyloid plaques found in the brains of Alzheimerandapos s disease patients PUBLIC HEALTH RELEVANCE Alzheimerandapos s disease is an age related non reversible brain disorder that develops over a period of years and the sixth leading cause of death in USA Recent studies estimates that there are million Americans of all ages have this disease In addition the number of people age and older with Alzheimerandapos s disease is estimated to reach million by There are three major markers in the brain that are associated with the process of Alzheimerandapos s disease Amyloid plaques neurofibrillary tangles and loss of connections between neurons Amyloid precursor protein APP which plays an important role in the development of Alzheimerandapos s disease is a membrane protein with several significant domains growth factor domain GFD copper binding domain CuBD Kunitz type protease inhibitor domain Most importantly the proteolysis of APP generates a neurotoxic A peptide which can affect neural functions and trigger cell death In addition A peptide can aggregate into small soluble oligomers eventually leading to the amyloid plaques observed in brains of patients who had Alzheimerandapos s disease Although the complete crystal structure of APP has not been solved most of its individual domains except A have been crystallized In this regard we propose new instrumentation i e named iCrystal and crystallization platforms that allows for rapid crystallization of APP and it components We envision that the iCrystal system can be applied to the crystallization of any biological macromolecule related to human diseases