My research program involves two areas:
(1) mass spectrometry.
We have two instruments: a Sciex API-III+ ESI/MS and a Perseptive DE-STR MALDI-TOF. We analyze post-translational modifications and are starting to work on proteome analysis. Proteome analysis is the identification of all the proteins in a given cell or biochemical preparation. Samples are analyzed on two dimensional gels, the gel is stained to locate the protein bands, which are then cut out and digested with proteases. The peptides are then analyzed by mass spectrometry and the information is used to do data base searches to identify the protein in the band. We are analyzing proteins of rat epidermis and cultures from those cells. In collaboration with Natalie Ahn, we are looking at K562 cells, and with David Norris from Denver Medical Center, we will be analyzing melanoma cells and epidermal cells from patients with acantholysis nigricans which have abnormal differentiation.
(2) epidermal differentiation:
The formation of squamous cells that slough from the surface of the skin is a tightly regulated process whereby living cells undergo major transformative events that destroy all cell organelles, remodel the cell membrane to form a thickened, cornified cell envelope, and convert the cell contents to a bag of aligned keratin filaments embedded in a matrix. The specific protein we are interested in is profilaggrin, a protein with several repeats that eventually become the matrix protein, filaggrin, as well as a calcium binding protein embedded in the cornified cell envelope. Processing to form the squamous cell is complex, involving several kinases, a phosphatase, and at least five proteases. In the squamous cell, the arginines in filaggrin are converted to citruline, which dissociates it from the keratin filaments. The filaggrin is then broken down to free amino acids (some of which are then deamidated to release ammonia); the resulting high salt concentrations provide the moisturizer of skin. We study the enzymes as well as characterize the specific modifications of profilaggrin, in order to understand the regulation of these events.
I attended U. Kansas and Washburn Univ. in Topeka for my undergraduate degree in Biology, taught High School Biology for two years in Topeka, then took time out to have kids (2 boys). I returned to school to get my Ph.D. from Dept of Biochem at U.W. in Seattle, where I worked for Ken Walsh. My initial project was working with a protease in sea urchin sperm, a project that produced many great stories, but not much scientific light. I shifted to work on epidermal differentiation and completed my Ph.D. a year later. I continued work in that system as a post-doc with Beverly Dale, also at U.W. I moved to Boulder to join the Chem and Biochem Dept in 1991.
1. Seyfried, C.E., Oleinik, O.E., Degen, J.L., Resing, K., and Morris, D.R. (1982) Purification, properties and regulation of the level of bovine s-adenosyl methionine decarboxylase during lymphocyte mitogenesis. Biochimica et Biophysica Acta. 716:169-177.
2. Titani, K., Sasagawa, T., Resing, K., and Walsh, K.A. (1982) A simple and rapid purification of commercial trypsin and chymotrypsin by reverse-phase high performance liquid chromatography. Analytical Biochemistry. 123:408-412.
3. Lonsdale-Eccles, J.D., Resing, K.A., Meek, R.L. and Dale, B.A. (1984) High molecular weight precursor of epidermal filaggrin and hypothesis for its tandem repeating structure. Biochemistry. 23:1239-1245.
4. Resing, K.A., Walsh, K.A. and Dale, B.A. (1984) Identification of two intermediates during processing of profilaggrin to filaggrin in neonatal mouse epidermis. Journal of Cell Biology. 99:1372-1378.
5. Resing, K.A.: Multiple domain structure and phosphorylation of profilaggrin. (1984) Doctoral dissertation, University of Washington.
6. Resing, K.A., Green, J.D., Walsh, K.A. (1985) 53,000 dalton esterase in dry Strongylocentrotus purpuratus sperm is from gonadal phagocytic cells. Developmental Biology 107:87-93.
7. Dale, B.A., Resing, K.A, Lonsdale-Eccles, J.D. (1985) Filaggrin, a keratin filament associated protein. Ann. N.Y. Acad. Sci. 455:330-342.
8. Resing, K.A., Peschon, J., Titani, K. (1985) Structure, function and evolution of epidermal proteases and their inhibitors. In: The Biological Role of Proteinases and Their Inhibitors in Skin. (Eds., Ogawa, H., Lazarus, G.S., & Hopsu-Havu, V.K.). Univ. of Tokyo Press, pp. 39-48.
9. Resing, K.A., Dale, B.A., Walsh, K.A. (1985) Multiple copies of phosphorylated filaggrin in epidermal profilaggrin demonstrated by analysis of tryptic peptides. Biochemistry. 24:4167-4175.
10. Resing, K.A., Scofield-Haugen, J., Mamrack, M.D., Dale, B.A. (1987) Use of filaggrin as a substrate for epidermal kinases and phosphatases. In: Processes in Cutaneous Epidermal Differentiation (Eds., Bernstein, I.A. and Hirone, T.). Praeger Scientific, pp. 165-186.
11. Dale, B.A., Gown, A.M., Fleckman, P., Kimball, J.R., Resing, K.A. (1987) Characterization of two monoclonal antibodies to human keratohyalin: reactivity with filaggrin and related proteins. J. Invest. Dermat. 88:306-313.
12. Haugen-Scofield, J., Resing, K.A., Dale, B.A. (1988) Purification of an epidermal phosphatase specific for filaggrin phosphorylated by casein kinase II. J. Invest. Dermat. 91:553-559.
13. Resing, K.A., Walsh, K.A., Haugen-Scofield, J., Dale, B.A. (1988) Identification of proteolytic cleavage sites in the conversion of profilaggrin to filaggrin in mammalian epidermis. J. Biol. Chem. 264:1837-1846.
14. Dale, B.A., Resing, K. A., Haydock, P.V., Fleckman, P., Fisher, C. and Holbrook, K.A. (1989) Intermediate Filament Associated Protein of Epidermis. In: The Biology of Wool and Hair (Eds., Rogers, G.E., Reis, P.J., Ward, K.A., and Marshall, R.C.). Chapman and Hall, Ltd. London, pp. 97-115.
15. Dale, B.A., Resing, K.A., and Haydock, P.V. (1991) Filaggrins. In: Cellular and Molecular Biology of Intermediate Filaments, (Eds., Goldman, R. and Steinert, P.M.). Plenum Publ. Corp, pp. 393-412.
16. Resing, K.A. and Dale, B.A. (1991) Proteins of Keratohyalin. In: Biochemistry and Physiology of the Skin. (Ed., Goldsmith, L.A.). Oxford University Press. Vol 1, pp 148-167.
17. Walsh, K.A., Erickson, L.H., Resing, K.A., and Johnson, R.S (1992). Electrospray-mass spectrometry, an emerging methodology for elucidating structure-function relationships of proteins. In Methods in Protein Sequence Analysis (Eds. Imahori K., Sakiyama F.) Plenum Press. pp. 143-148.
18. Dale, B.A., Presland, R., Resing, K.A., and Kam, E. (1993) Phenotypic expression and processing of filaggrin in epidermal differentiation. In Molecular Biology of the Skin. (ed. Darmon, M. & Blumenberg, M.). Academic Press pp. 79-106.
19. Kam, E., Resing, K.A., Lim, S., and Dale, B.A. (1993) Characterization of rat epidermal profilaggrin phosphatase as a type 2A protein phosphatase. J. of Cell Science. 106: 219-226.
20. Resing, K.A., Johnson, R.S., and Walsh, K.A. (1993) Characterization of protease processing sites during conversion of profilaggrin to filaggrin. Biochem. 32:10036-10045.
21. Resing, K.A., Al-Alawi, N., Blomquist, C., Fleckman, P., and Dale, B.A. (1993) Independent Regulation of two cytoplasmic processing stages of the intermediate filament associated protein filaggrin and role of Ca2+ in the second stage. J. Biol. Chem. 268:25139-25145.
22. Dale, B.A., Resing, K.A., and Presland, R.B. (1994) Keratohyalin granule proteins. In The Keratinocyte Handbook. (Eds. Leigh, I.M., Lane, E.B., Watt, F.M.) Cambridge University Press. pp. 323-350.
23. Gaudiano, G., Resing, K., Koch, T.H. (1994) Reaction of anthracycline antitumor drugs with reduced glutathione. Formation of aglycon conjugates. J. Am. Chem. Soc. 116:6537-6544
24. Resing, K.A., Mansour, S.J., Hermann, A.S., Johnson, R.S., Candia, J.M., Ahn, N.G. (1995) Determination of v mos catalyzed phosphorylation sites and auto-phosphorylation sites on MAP kinase kinase by ESI/MS. Biochemistry. 34:2610-2620.
25. Berger, S.J., Resing, K.A., Taylor, T.C., Melancon, P. (1995) Mass spectrometric analysis of ADP-ribosylation factors from bovine brain: identification and evidence for homogeneous myristoylation by the 14:0 fatty acid. Biochem. J., 311: 125-132.
26. Resing, K.A., Johnson, R.S., Walsh, K.A. (1995) Identification of twenty-one phosphorylation sites in the internal repeat of rat profilaggrin; precursor of an intermediate filament associated protein. Biochemistry, 34:9477-9487.
27. Resing, K.A., Craig Thulin, C., Whiting, K., Johnson, R.S., Al-Alawi, N., Mostad, S., and Walsh, K.A. (1995) Characterization of profilaggrin endoproteinase I; a regulated cytoplasmic protease of epidermis. J. Biol. Chem., 270:28193-28198.
28. Taatjes, D.J., Gaudiano, G., Resing, K.A., Koch, T.H. (1996) Alkylation of DNA by the antracycline, antitumor drugs adriamycin and daunomycin. J. of Med. Chem., 21: 4135-4138.
29. Louie, D., Resing, K.A., Ahn, N.G. (1996) Analysis of 40S ribosome proteins by mass spectrometry. J. Biol. Chem., 271:28189-28198.
30. Resing, K.A., Ahn, N.G. (1997) Analysis of protein phosphorylation. In: Methods of Enzymology, 283:29-44.
31. Ahn, N.G., Galasinski, S.G., Gloor, K.K., Lewis, T.S., Louie, D.F., Mansour, S.J., Nahreini, T.S., Resing, K.A., Shapiro, P.S., Whalen, A.M. (1997) Signal transduction through the MAP kinase pathway. NATO ASI Series. 102:147-151.
32. Resing, K.A., Ahn, N.G. (1998) Structural dynamics of MAPKK analyzed by deuterium exchange mass spectrometry. Biochemistry (in press).
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