cDNA and deduced amino acid sequence of human pulmonary surfactant-associated proteolipid SPL(Phe)

Journal Article · Mon Jun 01 00:00:00 EDT 1987 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.84.12.4007· OSTI ID:5445621

Glasser, S W; Korfhagen, T R; Weaver, T; Pilot-Matias, T; Fox, J L; Whitsett, J A

Hydrophobic surfactant-associated protein of M/sub r/ 6000-14,000 was isolated from either/ethanol or chloroform/methanol extracts of mammalian pulmonary surfactant. Automated Edman degradation in a gas-phase sequencer showed the major N-terminus of the human low molecular weight protein to be Phe-Pro-Ile-Pro-Leu-Pro-Try-Cys-Trp-Leu-Cys-Arg-Ala-Leu-. Because of the N-terminal phenylalanine, the surfactant protein was designated SPL(Phe). Antiserum generated against hydrophobic surfactant protein(s) from bovine pulmonary surfactant recognized protein of M/sub r/ 6000-14,000 in immunoblot analysis and was used to screen a lambdagt11 expression library constructed from adult human lung poly(A)/sup +/ RNA. This resulted in identification of a 1.4-kilobase cDNA clone that was shown to encode the N-terminus of the surfactant polypeptide SPL(Phe) (Phe-Pro-Ile-Pro-Leu-Pro-) within an open reading frame for a larger protein. Expression of a fused ..beta..-galactosidase-SPL (Phe) gene in Escherichia coli yielded an immunoreactive M/sub r/ 34,000 fusion peptide. Hybrid-arrested translation with the cDNA and immunoprecipitation of (/sup 35/S)methionine-labeled in vitro translation products of human poly(A)/sup +/ RNA with a surfactant polyclonal antibody resulted in identification of a M/sub r/ 40,000 precursor protein. Blot hybridization analysis of electrophoretically fractionated RNA from human lung detected a 2.0-kilobase RNA that was more abundant in adult lung than in fetal lung. These proteins, and specifically SPL(Phe), may therefore be useful for synthesis of replacement surfactants for treatment of hyaline membrane disease in newborn infants or of other surfactant-deficient states.

• Research Organization: Univ. of Cincinnati, OH
• OSTI ID: 5445621
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 84:12
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
LIPOPROTEINS
AMINO ACID SEQUENCE
DNA SEQUENCING
AGE DEPENDENCE
METHIONINE
PURIFICATION
RECOMBINANT DNA
RESPIRATORY SYSTEM DISEASES
SULFUR 35
SURFACTANTS
AMINO ACIDS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOXYLIC ACIDS
DAYS LIVING RADIOISOTOPES
DISEASES
DNA
DRUGS
EVEN-ODD NUCLEI
ISOTOPES
LIGHT NUCLEI
LIPIDS
LIPOTROPIC FACTORS
MOLECULAR STRUCTURE
NUCLEI
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PROTEINS
RADIOISOTOPES
STRUCTURAL CHEMICAL ANALYSIS
SULFUR ISOTOPES
550201* - Biochemistry- Tracer Techniques