Dissociation of surfactant protein B from canine surfactant large aggregates during formation of small surfactant aggregates by in vitro surface area cycling

1999 ◽  
Vol 1440 (1) ◽  
pp. 49-58 ◽  
Author(s):  
Kevin Inchley ◽  
Amanda Cockshutt ◽  
Ruud Veldhuizen ◽  
Fred Possmayer
Keyword(s):  
Surface Area ◽  
Surfactant Protein ◽  
Surfactant Aggregates ◽  
Surfactant Protein B ◽  
Protein B

scholarly journals Degradation of surfactant-associated protein B (SP-B) during in vitro conversion of large to small surfactant aggregates

1993 ◽  
Vol 295 (1) ◽  
pp. 141-147 ◽  
Author(s):  
R A W Veldhuizen ◽  
K Inchley ◽  
S A Hearn ◽  
J F Lewis ◽  
F Possmayer
Keyword(s):  
Surface Area ◽  
Sucrose Gradient ◽  
Gradient Centrifugation ◽  
Sucrose Gradient Centrifugation ◽  
Aggregate Fraction ◽  
Surfactant Aggregates ◽  
Morphological Studies ◽  
Surface Active ◽  
Protein B

Pulmonary surfactant obtained from lung lavages can be separated by differential centrifugation into two distinct subfractions known as large surfactant aggregates and small surfactant aggregates. The large-aggregate fraction is the precursor of the small-aggregate fraction. The ratio of the small non-surface-active to large surface-active surfactant aggregates increases after birth and in several types of lung injury. We have utilized an in vitro system, surface area cycling, to study the conversion of large into small aggregates. Small aggregates generated by surface area cycling were separated from large aggregates by centrifugation at 40,000 g for 15 min rather than by the normal sucrose gradient centrifugation. This new separation method was validated by morphological studies. Surface-tension-reducing activity of total surfactant extracts, as measured with a pulsating-bubble surfactometer, was impaired after surface area cycling. This impairment was related to the generation of small aggregates. Immunoblot analysis of large and small aggregates separated by sucrose gradient centrifugation revealed the presence of detectable amounts of surfactant-associated protein B (SP-B) in large aggregates but not in small aggregates. SP-A was detectable in both large and small aggregates. PAGE of cycled and non-cycled surfactant showed a reduction in SP-B after surface area cycling. We conclude that SP-B is degraded during the formation of small aggregates in vitro and that a change in surface area appears to be necessary for exposing SP-B to protease activity.

Surfactant protein B labelled with [99mTc(CO)3(H2O)3]+ retains biological activity in vitro

2001 ◽  
Vol 28 (3) ◽  
pp. 243-250 ◽  
Author(s):  
Anton Amann ◽  
Clemens Decristoforo ◽  
Ingo Ott ◽  
Martin Wenger ◽  
Dietmar Bader ◽  
...  
Keyword(s):  
Biological Activity ◽  
Surfactant Protein ◽  
Surfactant Protein B ◽  
Protein B

scholarly journals Surface film formation in vitro by infant and therapeutic surfactants: role of surfactant protein B

2014 ◽  
Vol 77 (2) ◽  
pp. 340-346 ◽  
Author(s):  
Olivier Danhaive ◽  
Cheryl Chapin ◽  
Hart Horneman ◽  
Paola E. Cogo ◽  
Philip L. Ballard
Keyword(s):  
Film Formation ◽  
Surface Film ◽  
Surfactant Protein ◽  
Surfactant Protein B ◽  
Protein B

scholarly journals Allele-specific N-glycosylation delays human surfactant protein B secretion in vitro and associates with decreased protein levels in vivo

2013 ◽  
Vol 74 (6) ◽  
pp. 646-651 ◽  
Author(s):  
Saija Taponen ◽  
Johanna M. Huusko ◽  
Ulla E. Petäjä-Repo ◽  
Reija Paananen ◽  
Susan H. Guttentag ◽  
...  
Keyword(s):  
Surfactant Protein ◽  
Protein Levels ◽  
Surfactant Protein B ◽  
Allele Specific ◽  
Protein B

scholarly journals Reversal of Surfactant Protein B Deficiency in Patient Specific Human Induced Pluripotent Stem Cell Derived Lung Organoids by Gene Therapy

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Sandra Lawrynowicz Leibel ◽  
Alicia Winquist ◽  
Irene Tseu ◽  
Jinxia Wang ◽  
Daochun Luo ◽  
...  
Keyword(s):  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Human Lung ◽  
Induced Pluripotent Stem Cell ◽  
Surfactant Protein ◽  
Patient Specific ◽  
Surfactant Protein B ◽  
Induced Pluripotent ◽  
Protein B

Abstract Surfactant protein B (SFTPB) deficiency is a fatal disease affecting newborn infants. Surfactant is produced by alveolar type II cells which can be differentiated in vitro from patient specific induced pluripotent stem cell (iPSC)-derived lung organoids. Here we show the differentiation of patient specific iPSCs derived from a patient with SFTPB deficiency into lung organoids with mesenchymal and epithelial cell populations from both the proximal and distal portions of the human lung. We alter the deficiency by infecting the SFTPB deficient iPSCs with a lentivirus carrying the wild type SFTPB gene. After differentiating the mutant and corrected cells into lung organoids, we show expression of SFTPB mRNA during endodermal and organoid differentiation but the protein product only after organoid differentiation. We also show the presence of normal lamellar bodies and the secretion of surfactant into the cell culture medium in the organoids of lentiviral infected cells. These findings suggest that a lethal lung disease can be targeted and corrected in a human lung organoid model in vitro.

Membrane Activity of (Cys48Ser) Lung Surfactant Protein B Increases with Dimerisation

2001 ◽  
Vol 382 (6) ◽  
pp. 933-939 ◽  
Author(s):  
Shahparak Zaltash ◽  
William J. Griffiths ◽  
David Beck ◽  
Chen-Xia Duan ◽  
Timothy E. Weaver ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Transgenic Mice ◽  
Ion Pairs ◽  
Lung Surfactant ◽  
Surfactant Protein ◽  
Cd Spectroscopy ◽  
Surfactant Protein B ◽  
Lung Surfactant Protein ◽  
Protein B

Abstract One of the possible functions of lung surfactant protein B (SPB), an hydrophobic membraneassociated saposinlike protein, is to reduce the alveolar surface tension by promoting insertion of phospholipids into the air/liquid interface of the lung. SPB is a covalent homodimer; Cys48 of two polypeptides form an intermolecular disulphide bond. In order to test whether dimerisation of SPB is important for surfactant function, transgenic mice which express (Cys48Ser) human SPB in a mouse SPB null background were generated. In previous studies (Cys48Ser)SPB showed a concentrationdependent in vitro activity, suggesting that it may form noncovalent dimers. Here (Cys48Ser)SPB isolated from bronchoalveolar lavage of transgenic mice was studied at different concentrations by circular dichroism (CD) spectroscopy, pulsating bubble surfactometry, mass spectrometry and reversedphase HPLC. The results indicate that (Cys48Ser)SPB, both in a phospholipid environment and in organic solvents, is largely monomeric and exhibits low activity at concentrations lower than 1 2 M, while at higher concentrations it forms noncovalent dimers, which are nearly functionally equivalent to native SPB in vitro. Furthermore, electrospray mass spectrometry showed that more dimers were found relative to the monomer when the polarity of the solvent was decreased, and when the concentration of SPB increased. (Cys48Ser)SPB also eluted earlier than native SPB in reversedphase HPLC. Taken together, these results indicate that a polar surface is buried upon dimerisation, thereby promoting formation of interchain ion pairs between Glu51 Arg52 and Glu51Arg52.

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