Blackburn Lab

About Us

Our lab focuses on the structure and function of oxidase and oxygenase metalloenzymes, particularly copper monooxygenases. We specialize in coordination chemistry and the biochemistry of copper, with particular focus in enzymes which influence neuroendocrine function, pathogen virulence, and cancer. Our biophysical areas of expertise include X-ray absorption spectroscopies (XAS, EXAFS, X-ray crystallography), spectroscopy (EPR, UV-Vis, fluorescence, FTIR), and rapid kinetics (RFQ, stopped-flow, quench-flow microvolume).

People

Professor Ninian Blackburn

Ninian's email

Ph.D., 1975, University of Dundee, Scotland

Specialties:

Biochemistry of metalloproteins, particularly on the role of copper in neuroendocrine function and investigation of metal ion homeostasis
Advanced X-ray spectroscopies (XAS, EXAFS, XES)
Coordination chemistry
Electron paramagnetic resonance (EPR)
Fourier-transform infrared (FTIR) spectroscopy

Postdoctoral Researcher, Renee Arias, PhD

Dr. Renee Arias

Renee's email

Ph.D., 2018, California Institute of Technology, Pasadena, CA

B.S., 2012, University of Oregon, Eugene, OR

Specialties:

Investigation of the structure and function of metalloproteins, with emphasis on FeS and Cu proteins
X-ray crystallography
Anaerobic protein purification (liquid chromatography, ion exchange, metal reconstitution)
Rapid kinetics and analysis: stopped-flow (SF), microvolume quench-flow (QFM), HPLC
X-ray spectroscopy (XAS, HERFD)

Postdoctoral Researcher, Katie Rush, PhD

Dr. Katie Rush

Katie's email

Ph.D., 2018, University of Michigan, Ann Arbor, MI

B.S., 2013, University of Tennessee, Knoxville, TN

Specialties:

Biochemical investigation of metalloproteins, with emphasis on Hg, FeS, and Cu systems
X-ray spectroscopies (XAS, HERFD)
Rapid kinetics: rapid freeze-quench (RFQ)
Electron paramagnetic resonance (EPR)
Protein purification (liquid chromatography, ion exchange)

Dr. Rush is co-advised by Professor Blackburn at OHSU, and Professor Kelly Chacón at Reed College

Katherine Alwan

Katherine's email

M.S., 2015, Oregon Health and Science University, Portland, OR

B.A., 2014, Skidmore College, Saratoga Springs, NY

Specialties:

Use of rationally designed protein scaffolds to investigate peptidylglycine alpha-hydroxylating monooxygenase active site function
X-ray spectroscopies (XAS)
Electron paramagnetic resonance (EPR)
Fourier transform infrared (FTIR) spectroscopy
Protein purification
Kinetics evaluation (SigmaPlot)

Evan Welch

Evan's email

B.A., 2017, Reed College, Portland, OR

Specialties

Investigation of peptidylglycine alpha-amidating monooxygenase function using biochemical and structural methods
Protein purification (liquid chromatography, ion exchange, metal reconstitution)
Fourier transform infrared (FTIR) spectroscopy
Rapid kinetics and analysis: microvolume quench flow (QFM), HPLC
Kinetics evaluation and programming: DynaFit, SigmaPlot, R, Python

Ben Gambill

B.S., 2014, Oregon State University, Corvallis, OR

Ben's email

Specialties:

Mammalian cloning and cell culture
General lab maintenance

Research Projects

Metallochaperone scaffold proteins as models to investigate copper monooxygenase structure and function.

Peptidylglycine alpha-amidating monooxygenase with spectrum

Investigation of peptidylglycine alpha-amidating monooxygenase chemical structure and reactivity.

Research Specialties

Advanced X-ray spectroscopies: X-ray absorption (XAS) and Extended X-ray Absorption Fine Structure (EXAFS)
X-ray crystallography
Fourier transform infrared (FTIR) spectroscopy
UV-visible spectroscopy (UV-vis)
Electron paramagnetic resonance (EPR)
Fast kinetics: stopped-flow (SF), rapid freeze-quench (RFQ), quench flow microvolume (QFM)

Recent Publications

pH-Induced Binding of the Axial Ligand in an Engineered CuA Site Favors the πu State.

Morgada MN, Emiliani F, Chacón KN, Álvarez-Paggi D, Murgida DH, Blackburn NJ, Abriata LA, Vila AJ. Inorg Chem. 2019 Dec 2;58(23):15687-15691. doi: 10.1021/acs.inorgchem.9b01868. Epub 2019 Nov 11. PubMed PMID: 31710470.

Catalytic M Center of Copper Monooxygenases Probed by Rational Design. Effects of Selenomethionine and Histidine Substitution on Structure and Reactivity.

Alwan KB, Welch EF, Blackburn NJ. Biochemistry. 2019 Nov 5;58(44):4436-4446. doi: 10.1021/acs.biochem.9b00823. Epub 2019 Oct 28. PMID: 31626532

The binuclear cluster of [FeFe] hydrogenase is formed with sulfur donated by cysteine of an [Fe(Cys)(CO)₂(CN)] organometallic precursor.

Rao G, Pattenaude SA, Alwan K, Blackburn NJ, Britt RD, Rauchfuss TB. Proc Natl Acad Sci U S A. 2019 Oct 15;116(42):20850-20855. doi: 10.1073/pnas.1913324116. Epub 2019 Sep 30. PubMed PMID: 31570604; PubMed Central PMCID: PMC6800375.

Incorporation of Ni²⁺, Co²⁺, and Selenocysteine into the Auxiliary Fe-S Cluster of the Radical SAM Enzyme HydG.

Rao G, Alwan KB, Blackburn NJ, Britt RD. Inorg Chem. 2019 Oct 7;58(19):12601-12608. doi: 10.1021/acs.inorgchem.9b01293. Epub 2019 Sep 20. PubMed PMID: 31539235; PubMed Central PMCID: PMC6984664.

Rational Design of a Histidine-Methionine Site Modeling the M-Center of Copper Monooxygenases in a Small Metallochaperone Scaffold.

Alwan KB, Welch EF, Arias RJ, Gambill BF, Blackburn NJ. Biochemistry. 2019 Jul 16;58(28):3097-3108. doi: 10.1021/acs.biochem.9b00312. Epub 2019 Jun 27. PMID: 31243953

Trapping intermediates in metal transfer reactions of the CusCBAF export pump of Escherichia coli.

Chacón KN, Perkins J, Mathe Z, Alwan K, Ho EN, Ucisik MN, Merz KM, Blackburn NJ. Commun Biol. 2018 Nov 14;1:192. doi: 10.1038/s42003-018-0181-9. eCollection 2018. PMID: 30456313

Substrate-Induced Carbon Monoxide Reactivity Suggests Multiple Enzyme Conformations at the Catalytic Copper M-Center of Peptidylglycine Monooxygenase.

Kline CD, Blackburn NJ. Biochemistry. 2016 Dec 6;55(48):6652-6661. Epub 2016 Nov 22. PMID: 27933800

pH-regulated metal-ligand switching in the HM loop of ATP7A: a new paradigm for metal transfer chemistry.

Kline CD, Gambill BF, Mayfield M, Lutsenko S, Blackburn NJ. Metallomics. 2016 Aug 1;8(8):729-33. doi: 10.1039/c6mt00062b. PMID: 27242196

Stopped-Flow Studies of the Reduction of the Copper Centers Suggest a Bifurcated Electron Transfer Pathway in Peptidylglycine Monooxygenase.

Chauhan S, Hosseinzadeh P, Lu Y, Blackburn NJ. Biochemistry. 2016 Apr 5;55(13):2008-21. doi: 10.1021/acs.biochem.6b00061. Epub 2016 Mar 23. PMID: 26982589

Kβ Valence to Core X-ray Emission Studies of Cu(I) Binding Proteins with Mixed Methionine - Histidine Coordination. Relevance to the Reactivity of the M- and H-sites of Peptidylglycine Monooxygenase.

Martin-Diaconescu V, Chacón KN, Delgado-Jaime MU, Sokaras D, Weng TC, DeBeer S, Blackburn NJ. Inorg Chem. 2016 Apr 4;55(7):3431-9. doi: 10.1021/acs.inorgchem.5b02842. Epub 2016 Mar 11. PMID: 26965786