Measurement of human surfactant protein-B turnover in vivo from tracheal aspirates using targeted proteomics

Daniela M. Tomazela, Bruce W. Patterson, Elizabeth Hanson, Kimberly L. Sponce, Tiffany B. Kanion, David H. Salinger, Paolo Vicini, Hugh Barret, Hillary B. Heins, F. Sessions Cole, Aaron Hamvas, Michael J. MacCoss

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18 Scopus citations

Abstract

We describe a method to measure protein synthesis and catabolism in humans without prior purification and use the method to measure the turnover of surfactant protein-B (SP-B). SP-B, a lung-specific, hydrophobic protein essential for fetal-neonatal respiratory transition, is present in only picomolar quantities in tracheal aspirate samples and difficult to isolate for dynamic turnover studies using traditional in vivo tracer techniques. Using infusion of [5,5,5-2H3] leucine and a targeted proteomics method, we measured both the quantity and kinetics of SP-B tryptic peptides in tracheal aspirate samples of symptomatic newborn infants. The fractional synthetic rate (FSR) of SP-B measured using the most abundant proteolytic fragment, a 10 amino acid peptide from the carboxy-terminus of proSP-B (SPTGEWLPR), from the circulating leucine pool was 0.035 ± 0.005 h- 1, and the fractional catabolic rate was 0.044 ± 0.003 h -1. This technique permits high-throughput and sensitive measurement of turnover of low abundance proteins with minimal sample preparation.

Original languageEnglish
Pages (from-to)2561-2567
Number of pages7
JournalAnalytical Chemistry
Volume82
Issue number6
DOIs
StatePublished - Mar 15 2010

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    Tomazela, D. M., Patterson, B. W., Hanson, E., Sponce, K. L., Kanion, T. B., Salinger, D. H., Vicini, P., Barret, H., Heins, H. B., Sessions Cole, F., Hamvas, A., & MacCoss, M. J. (2010). Measurement of human surfactant protein-B turnover in vivo from tracheal aspirates using targeted proteomics. Analytical Chemistry, 82(6), 2561-2567. https://doi.org/10.1021/ac1001433