Biochemistry Core

National Heart, Lung, And Blood Institute

Investigators

Linked publications & trials

Abstract

1. Core utilization and accomplishments - A total of 53 people from 28 laboratories have used for the facility services and instruments. - Support investigators by implementing methods for 1) mass measurement of intact proteins and organic compounds (a total of 2,570 samples from 22 laboratories), and 2) separation and quantification of proteins or metabolites by HPLC (a total of 1,346 samples from 6 laboratories). - Applying a new method for the quantification of small organic compounds by LC-MS and developing of a new sample preparation method to speed up data collection. 2. Core publications Liu, Y., et al., The PPR domain of mitochondrial RNA polymerase is a ribonuclease required for mtDNA replication. Nat. Cell Biol. 24, 757-765 (2022) Sydney O., et al., Wathcing liquid droplets of TDP-43ctD age by Raman spectroscopy. J.Biol.Chem. 298, 101538 (2022) Strickland, M., et al., Tsg101/ESCRT-1 recruitment regulated by the dual binding modes of K63-linked diubiquitin. Structure 30, 289-299 (2022) Pasapera, A., et al., MARK2 regulates directed cell migration through modulation of myosin II contractility and focal adhesion organization. Curr.Biol 32, 2704-2718 (2022) Nyenhuis, D., et al., Incorporation of residual chemical shift anisotropy into the treatment of 15N pseudocontact shifts for structural refinement. J. Magn. Reson. 340, 107213 (2022) McGlinchey, R., et al., The N terminus of a-synuclein dictates fibril formation. Proc Natl Acad Sci U S A 118 e2023487118 (2021) Chen, J., et al., -tubulin tail modifications regulate microtubule stability through selective effector recruitment, not changes in intrinsic polymer dynamics. Dev. Cell 56, 1-13 (2021) Lyu, J., et al., The short-chain fatty acid acetate modulates epithelial-to-mesenchymal transition. Mol.Biol.Cell 33, No.8 (2022) Zhu, J., et al., Kruppel-like factor 1-GATA1 fusion protein improves the sickle cell disease phenotype both in vitro and in vivo. Blood, In press (2022) Uchida, N., et al., Preclinical evaluation for engraftment of CD34+ cells gene-edited at the sickle cell disease locus in xenograft mouse and non-human primate models. Cell Rep. Med. 2, 1-11 (2021) 3. New instrumentation and new services System upgrade for Arthur (TOF-LC/MS) - Replaced old DAD, binary pump, and autosampler - Upgraded the controller PC with the latest version of data acquisition program Upgraded the workstation for LC/MS data analysis with the latest version of the analysis program Full set of HPLC systems (Property transferred from Dr. Carole Bewley in NIDDK): Backup for the HPLC in Bldg. 10 - Quaternary pump, autosampler with thermostat, fraction collector-prep scale, DAD, FLD, and RID Identification of small molecules using LC-MS/MS with the METLIN Gen 2 database that has MS/MS data on 860,000 molecular standards (metabolites, drug compounds, etc.). - Simultaneous identification and detection of N-arachidonoyl glycine and arachidonic acid in plasma by QTOF-LC/MS (Dr. Michael Sacks lab) - PK test of Metformin and propranolol (IS) in monkey plasma (Dr. Cynthia Dunbars lab) - Identification of cADP-ribose, ADP-ribose, ADP-ribose-phosphate, ADP-ribosyl arginine (Dr. Joel Moss lab) - Identification of unknown lipids bound on co-purified protein (Dr. Jiansen Jiang lab) 4. FY202 budget The budget spent 99.9% as of August 23, 2022. - Maintenance contract for LC/MS---- 85% - Solvents and chemicals------------ 10% - General consumables---------------- 5%