Craig A Aspinwall
Professor, BIO5 Institute
Professor, Biomedical Engineering
Professor, Chemistry and Biochemistry-Sci
Primary Department
(520) 621-6338
Research Interest
Craig A. Aspinwall, PhD, is an Associate Professor of Chemistry and Biochemistry at the University of Arizona. Dr. Aspinwall’s research is focused on the development of novel technology that facilitates the investigation of the molecular underpinnings of disease states. His work encompasses a broad range of scientific disciplines and allows complex biochemical problems to be studied with an increasing level of molecular detail. Dr. Aspinwall has published over 40 original research papers and maintains active collaborations with several international investigators. His research has been funded by the National Institutes of Health, the National Science Foundation, the Arizona Biomedical Research Corporation, and other organizations. He is actively involved in mentoring and education of students and young scientists.

Publications

Johnson, G. M., Chozinski, T. J., Gallagher, E. S., Aspinwall, C. A., & Miranda, K. M. (2014). Glutathione sulfinamide serves as a selective, endogenous biomarker for nitroxyl after exposure to therapeutic levels of donors. Free radical biology & medicine, 76, 299-307.

Nitroxyl (HNO) donors exhibit promising pharmacological characteristics for treatment of cardiovascular disorders, cancer, and alcoholism. However, whether HNO also serves as an endogenous signaling agent is currently unknown, largely because of the inability to selectively and sensitively detect HNO in a cellular environment. Although a number of methods to detect HNO have been developed recently, sensitivity and selectivity against other nitrogen oxides or biological reductants remain problematic. To improve selectivity, the electrophilic nature of HNO has been harnessed to generate modifications of thiols and phosphines that are unique to HNO, especially compared to nitric oxide (NO). Given high bioavailability, glutathione (GSH) is expected to be a major target of HNO. As a result, the putative selective product glutathione sulfinamide (GS(O)NH2) may serve as a high-yield biomarker of HNO production. In this work, the formation of GS(O)NH2 after exposure to HNO donors was investigated. Fluorescent labeling followed by separation and detection using capillary zone electrophoresis with laser-induced fluorescence allowed quantitation of GS(O)NH2 with nanomolar sensitivity, even in the presence of GSH and derivatives. Formation of GS(O)NH2 was found to occur exclusively upon exposure of GSH to HNO donors, thus confirming selectivity. GS(O)NH2 was detected in the lysate of cells treated with low-micromolar concentrations of HNO donors, verifying that this species has sufficient stability to server as a biomarker of HNO. Additionally, the concentration-dependent formation of GS(O)NH2 in cells treated with an HNO donor suggests that the concentration of GS(O)NH2 can be correlated to intracellular levels of HNO.

Aspinwall, C., Hapuarachchi, S., Premeau, S. P., & Aspinwall, C. A. (2006). High-speed capillary zone electrophoresis with online photolytic optical injection. Analytical chemistry, 78(11).

We report an online, optical injection interface for capillary zone electrophoresis (CZE) based upon photophysical activation of a caged, fluorogenic label covalently attached to the target analyte. This injection interface allows online analysis of biomolecular systems with high temporal resolution and high sensitivity. Samples are injected onto the separation capillary by photolysis of a caged-fluorescein label using the 351-364 nm irradiation of an Ar+ laser. Following injection, the sample is separated and detected via laser-induced fluorescence detection at 488 nm. Detection limits for online analysis of arginine, glutamic acid, and aspartic acid were less than 1 nM with separation times less than 5 s and separation efficiencies exceeding 1,000,000 plates/m. Rapid injection of proteins was demonstrated with migration times less than 500 ms and 0.5 nM detection limits. Online monitoring was performed with response times less than 20 s, suggesting the feasibility of this approach for online, in vivo analysis for a range of biologically relevant analytes.

Jung, S., Aspinwall, C. A., & Kennedy, R. T. (1999). Detection of multiple patterns of oscillatory oxygen consumption in single mouse islets of Langerhans. Biochemical and Biophysical Research Communications, 259(2), 331-335.

PMID: 10362508;Abstract:

A novel oxygen microsensor was used to measure oxygen levels in single mouse islets as a function of glucose concentration. Oxygen consumption of individual islets was 5.99 ± 1.17, 9.21 ± 2.15, and 12.22 ± 2.16 pmol/min at 3, 10, and 20 mM glucose, respectively (mean ± SEM, n = 10). Consumption of oxygen was islet-size dependent as larger islets consumed more oxygen than smaller islets but smaller islets consumed more oxygen per unit volume than larger islets. Elevating glucose levels from 3 to 10 mM induced pronounced fast oscillations in oxygen level (period of 12.1 ± 1.7 s, n = 6) superimposed on top of large slow oscillations (period of 3.3 ± 0.6 min, n = 6). The fast oscillations could be completely abolished by treatment with the L-type Ca2+-channel blocker nifedipine (40 μM) with a lesser effect on slow oscillations. Slow oscillations were almost completely dependent upon extracellular Ca2+. The oxygen patterns closely mimic those that have previously been reported for intracellular Ca2+ levels and are suggestive of an important role for Ca2+ in amplifying metabolic oscillations.

Kennedy, R. T., Huang, L., & Aspinwall, C. A. (1996). Extracellular pH is required for rapid release of insulin from Zn - insulin precipitates in β-cell secretory vesicles during exocytosis. Journal of the American Chemical Society, 118(7), 1795-1796.
Aspinwall, C., Cheng, Z., D'Ambruoso, G. D., & Aspinwall, C. A. (2006). Stabilized porous phospholipid nanoshells. Langmuir : the ACS journal of surfaces and colloids, 22(23).

Chemically stabilized, porous phospholipid nanoshells (PPNs) were prepared via copolymerization of reactive monomers with unilamellar bis-Sorbyl phosphatidylcholine vesicles. The resulting PPN vesicular assemblies possess a highly porous membrane structure that allows passage of small molecules, which can react with encapsulated proteins and reporters. The unique combination of membrane stability and porosity will prove useful for preparing nanometer-sized sensor, container, and reactor platforms stable in harsh chemical and biological environments.