The role of charged amphipathic helices in the structure and function of surfactant protein B.
Surfactant protein B (SP-B) is essential for normal lung surfactant function. Theoretical models predict that the disulfide cross-linked, N- and C-terminal domains of SP-B fold as charged amphipathic helices, and suggest that these adjacent helices participate in critical surfactant activities. This hypothesis is tested using a disulfide-linked construct (Mini-B) based on the primary sequences of the N- and C-terminal domains. Consistent with theoretical predictions of the full-length protein, both isotope-enhanced Fourier transform infrared (FTIR) spectroscopy and molecular modeling confirm the presence of charged amphipathic alpha-helices in Mini-B. Similar to that observed with native SP-B, Mini-B in model surfactant lipid mixtures exhibits marked in vitro activity, with spread films showing near-zero minimum surface tensions during cycling using captive bubble surfactometry. In vivo, Mini-B shows oxygenation and dynamic compliance that compare favorably with that of full-length SP-B. Mini-B variants (i.e. reduced disulfides or cationic residues replaced by uncharged residues) or Mini-B fragments (i.e. unlinked N- and C-terminal domains) produced greatly attenuated in vivo and in vitro surfactant properties. Hence, the combination of structure and charge for the amphipathic alpha-helical N- and C-terminal domains are key to SP-B function.
Waring
A J
AJ
Department of Medicine, Division of Infectious Diseases, UCLA School of Medicine, Center for Health Sciences, Los Angeles, CA 90095, USA. awaring@ucla.edu
Walther
F J
FJ
Gordon
L M
LM
Hernandez-Juviel
J M
JM
Hong
T
T
Sherman
M A
MA
Alonso
C
C
Alig
T
T
Braun
A
A
Bacon
D
D
Zasadzinski
J A
JA
D000595:Amino Acid Sequence; D000818:Animals; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000494:pharmacology*; D011498:Protein Precursors / Q000737:chemistry*; D017433:Protein Structure, Secondary; D011510:Proteolipids / Q000737:chemistry*; D011663:Pulmonary Surfactants / Q000138:chemical synthesis / Q000494:pharmacology*; D051381:Rats; D017550:Spectroscopy, Fourier Transform Infrared; D013501:Surface-Active Agents / Q000138:chemical synthesis / Q000494:pharmacology*
D010455:Peptides; D011498:Protein Precursors; D011510:Proteolipids; D011663:Pulmonary Surfactants; D013501:Surface-Active Agents; C105511:surfactant protein B propeptide