Quantitation of elastin through measurement of its pentapeptide content |
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| Authors: | L B Sandberg T B Wolt J G Leslie |
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| Affiliation: | 1. Department of Chemistry, Centre for Advanced Studies in Chemistry, Panjab University, Sector-14, Chandigarh 160014, India;2. Central Scientific Instruments Organisation (CSIR-CSIO), Sector 30 C, Chandigarh 160030, India;3. Department of Applied Sciences, University Institute of Engineering & Technology, Panjab University, Sector-25, Chandigarh 160014, India;1. Department of Public Health and Infectious Diseases, Sapienza University of Rome, P.le Aldo Moro 5, 00185 Rome, Italy;2. Laboratory of Industrial Toxicology, Department of Clinical and Experimental Medicine, University of Parma, via Gramsci 14, I-43126 Parma, Italy;1. Center for Pharmacy and Experimental Therapeutics, University of Georgia College of Pharmacy, Augusta, GA, USA;2. Charlie Norwood VA Medical Center, Augusta, GA, USA;3. Department of Pharmacology and Toxicology, Faculty of Pharmacy, Mansoura University, Egypt;4. Division of Endocrinology, Diabetes, and Metabolism, Pennsylvania State University College of Medicine, Hershey, PA, USA;5. Department of Medicine, Pennsylvania State University College of Medicine, Hershey, PA, USA;6. Vascular Biology Center, Department of Pharmacology and Toxicology, Augusta University, Augusta, GA, USA |
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| Abstract: | Digestion of insoluble porcine elastin with thermolysin produces a number of discrete small peptides. That present in highest concentration is the pentapeptide valyl-glycyl-valyl-prolyl-glycine (VGVPG) derived from the portion of the polymer containing extensive repeats of this sequence. Among eukaryotes, this sequence appears to be found only in elastin and its precursor tropoelastin. In the pig this is represented by peptide W4 of a tropoelastin tryptic digest (Sandberg, L.B., et al. Path. Biol. 33, 266-274, 1985). Quantitation of this peptide by HPLC separation, monitoring its absorption at 212 nm, offers a simple reliable means of measuring purified insoluble elastin as well as non-purified elastin in fat-free tissue samples. Digestion times and incubation temperatures are discussed. The method is sensitive enough to accurately quantitate elastin at the 2 to 3 microgram level. |
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