Date of Award


Degree Name



College of Science

Type of Degree


Document Type


First Advisor

William Price

Second Advisor

Leslie Frost

Third Advisor

Seth Bush


Many human disorders are associated with the malfunction of smooth muscle tissue, or are related to the capabilities of its proper function—asthma, glaucoma, renal inefficiency, hypertension, and cardiovascular disease. Dysfunctional proteins are frequently implicated as the source of such disorders. As the second highest cause of death in the United States, the epidemic of cardiovascular disease makes the study of smooth muscle of utmost concern.

The capabilities of proteomics and mass spectrometry allow the entire proteome complement of a cell or tissue type to be analyzed at once. This investigation employs such techniques in an effort to better understand the mechanisms of signal transduction for the contraction of smooth muscle tissue. The differential phosphorylation of proteins between resting and contracted tissue was targeted, specifically, because phosphorylation is the primary means of communication and activity modification of proteins. Much of what is known about the regulation of signal transduction in smooth muscle contraction is conducted through the activation of myosin light chain kinase (MLCK) and/or myosin light chain phosphatase (MLCP).

Protocols of analysis using phosphoproteomic techniques and MALDI-TOF MS were developed for the purposes of this investigation. The localization of the more abundant proteins actin, myosin heavy chain (MHC), and tropomyosin was determined on a 2D gel. Tropomyosin was found to be phosphorylated in phorbol-contracted tissue, indicating that this protein may play a larger role in contractile regulation than originally thought. The function of such thin-filament associated proteins should be investigated more thoroughly. A thorough understanding of contraction in normal tissue will provide the base for which dysfunctions can be compared. Continued improvements in methodology will aid the advancement of this research.




Smooth muscle -- Contraction.