Subject(s)
Cost of Illness , Diabetes Mellitus, Type 2/prevention & control , Patient Education as Topic/organization & administration , Primary Prevention/organization & administration , Telemedicine/organization & administration , Algorithms , Diabetes Mellitus, Type 2/economics , Diabetes Mellitus, Type 2/etiology , Exercise Therapy , Florida , Health Expenditures/statistics & numerical data , Home Care Services/organization & administration , Humans , Life Style , Nurses/organization & administration , Nutritional Sciences/education , Obesity/complications , Obesity/prevention & control , Prediabetic State/complications , Prediabetic State/prevention & control , Risk Assessment , Social Support , United States , United States Department of Veterans AffairsABSTRACT
Partially purified transducin was resolved using two-dimensional gel electrophoresis (2-DE). Peptide mass fingerprinting of several different spots believed to correspond to the 37 kDa beta-subunit of transducin (T(beta)) was performed. Spots were excised and proteolyzed using modified trypsin. Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) was performed on the peptide mixture resulting from each spot. As many as six spots with different pI, ranging from 5.2 to 6.1, were observed when separated using 2-DE. MALDI peptide mass fingerprinting determined with high probability that all of the spots were the same gene product, guanine nucleotide-binding protein G(I)/G(S)/G(T) beta-subunit 1 (GNB1; T(beta1)). This suggested that post-translational modification was responsible for the differences in pI. Phosphorylation experiments showed that at least one T(beta1) spot was phosphorylated in vitro with [gamma-(32)P]ATP by an endogenous kinase. Treatment of T(beta) with alkaline phosphatase caused a large change in the spot pattern of T(beta), suggesting that phosphorylated T(beta) is a substrate for alkaline phosphatase. We conclude that T(beta1) constitutes over 99% of the T(beta) expressed in bovine rod outer segments and displays structural heterogeneity that is due to post-translational modification. We also conclude that some, but not all, of the heterogeneity observed is due to phosphorylation of Tb1.