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


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-(2)H(3)] 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.

SUBMITTER: Tomazela DM 

PROVIDER: S-EPMC2843406 | biostudies-literature | 2010 Mar

REPOSITORIES: biostudies-literature

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

Tomazela Daniela M DM   Patterson Bruce W BW   Hanson Elizabeth E   Spence Kimberly L KL   Kanion Tiffany B TB   Salinger David H DH   Vicini Paolo P   Barret Hugh H   Heins Hillary B HB   Cole F Sessions FS   Hamvas Aaron A   MacCoss Michael J MJ  

Analytical chemistry 20100301 6


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-(2)H(3)] leucine and a targeted pr  ...[more]

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