Matthew Amato Email & Phone Number

Dallas, Texas, United States

Rochester, Minnesota Area

Post-baccalaureate Research Education Program (PREP) engaging in biomedical research.Project Title: Investigating the dynamics of nucleosomes reconstituted with histone charge variants modeling a succinyllysine post-translational modification in the context of paraganglioma • Utilize a T7 RNA polymerase in vitro transcription assay to interrogate changes in single nucleosome stability induced by change-of-charge histone H4 mutants modeling succinyllysine histone modificationHypothesis: histone succinylation destabilizes nucleosomes o Utilizing experimental techniques  Reference mapping RNA transcripts by electrophoresis-based regression analysis  Generation of a heterogenous pool of RNA molecules transcribed in vitro Fixed position nucleosome reconstitution with a DNA template amenable to an in vitro transcription assay Molecular cloning a T7 promoter DNA template harboring a single histone octamer localization sequence with elements amenable to an in vitro transcription assay  Assembly of histone octamers by chaotropic dialysis Site-directed mutagenesis generation of histone H4 change-of-charge mutants Disclaimer: All statements expressed on this profile are my own and do not reflect the views of the Mayo Clinic*

I conducted biochemical research under the direction of Dr. Erik Yukl. The primary focus of the laboratory was to understand transition metal homeostasis in model gram negative organism Paracoccus Denitrificans. My contribution to the lab was in characterizing metal binding in a putative nickel ABC transporter. In this project I employed techniques in molecular cloning, FPLC based protein purification, and analytically sensitive biophysical assays.

Dallas/Fort Worth Area

Conducting biochemical research under the direction of Dr. Vincent Tagliabracci, I worked to elucidate the catalytic mechanism of HopBF1, a non-conical protein kinase. I employed a three step FPLC-based protein purification to produce HopBF1 in working quantities and then used a simplified purification procedure to generate 12 point mutants of HopBF1. The 12 point mutants were constructed to test for residues involved in catalysis. Performing a phosphorylation assay incubating HopBF1 with a putative substrate, utilizing radio-labeled ATP I measure catalytic activity of the mutants. I helped demonstrate that although HopBF1 shares marginal homology with classic protein kinases it still proceeds through a conical catalytic mechanism.