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"SP-G", a putative new surfactant protein--tissue localization and 3D structure.

Rausch F, Schicht M, Paulsen F, Ngueya I, Bräuer L, Brandt W - PLoS ONE (2012)

Bottom Line: In this work, computational chemistry and molecular-biological methods were combined to localize and characterize SP-G.With the help of a protein structure model, specific antibodies were obtained which allowed the detection of SP-G not only on mRNA but also on protein level.This includes also the possibility of interactions with lipid systems and with that, a potential surface-regulatory feature of SP-G.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioorganic Chemistry, Leibniz Institute of Plant Biochemistry, Halle, Germany.

ABSTRACT
Surfactant proteins (SP) are well known from human lung. These proteins assist the formation of a monolayer of surface-active phospholipids at the liquid-air interface of the alveolar lining, play a major role in lowering the surface tension of interfaces, and have functions in innate and adaptive immune defense. During recent years it became obvious that SPs are also part of other tissues and fluids such as tear fluid, gingiva, saliva, the nasolacrimal system, and kidney. Recently, a putative new surfactant protein (SFTA2 or SP-G) was identified, which has no sequence or structural identity to the already know surfactant proteins. In this work, computational chemistry and molecular-biological methods were combined to localize and characterize SP-G. With the help of a protein structure model, specific antibodies were obtained which allowed the detection of SP-G not only on mRNA but also on protein level. The localization of this protein in different human tissues, sequence based prediction tools for posttranslational modifications and molecular dynamic simulations reveal that SP-G has physicochemical properties similar to the already known surfactant proteins B and C. This includes also the possibility of interactions with lipid systems and with that, a potential surface-regulatory feature of SP-G. In conclusion, the results indicate SP-G as a new surfactant protein which represents an until now unknown surfactant protein class.

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Related in: MedlinePlus

Antibody test by Western blot.Analyzed was A) lung tissue which is used as positive control for surfactant proteins; B) recombinantly synthesized SP-G protein (not purified) at 28°C (1) and 37°C (2), arrows indicate positive evidence of the surfactant protein G. The proteins extracted from the lung tissue and separated by 15% SDS-PAGE under reducing conditions show distinct bands for SP-G at the theoretically expected molecular weights of 11, 20 and 30 kDA [A]. In case of recombinantly expressed SP-G protein, the antibody detects a distinct band at 11 kDa [B].
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pone-0047789-g007: Antibody test by Western blot.Analyzed was A) lung tissue which is used as positive control for surfactant proteins; B) recombinantly synthesized SP-G protein (not purified) at 28°C (1) and 37°C (2), arrows indicate positive evidence of the surfactant protein G. The proteins extracted from the lung tissue and separated by 15% SDS-PAGE under reducing conditions show distinct bands for SP-G at the theoretically expected molecular weights of 11, 20 and 30 kDA [A]. In case of recombinantly expressed SP-G protein, the antibody detects a distinct band at 11 kDa [B].

Mentions: The specificity of the resulting antibody was tested with protein from lung tissue (30 µg) and with the recombinantly synthesized SP-G protein (not purified, 30 µg) (Figure 7). The purified antibody shows distinct protein bands in lung for SP-G at 11 kDa, 20 kDa and 30 kDa and a distinct protein band for recombinantly synthesized SP-G at about 12 kDa. We used lung tissue as specific positive control for surfactant proteins.


"SP-G", a putative new surfactant protein--tissue localization and 3D structure.

Rausch F, Schicht M, Paulsen F, Ngueya I, Bräuer L, Brandt W - PLoS ONE (2012)

Antibody test by Western blot.Analyzed was A) lung tissue which is used as positive control for surfactant proteins; B) recombinantly synthesized SP-G protein (not purified) at 28°C (1) and 37°C (2), arrows indicate positive evidence of the surfactant protein G. The proteins extracted from the lung tissue and separated by 15% SDS-PAGE under reducing conditions show distinct bands for SP-G at the theoretically expected molecular weights of 11, 20 and 30 kDA [A]. In case of recombinantly expressed SP-G protein, the antibody detects a distinct band at 11 kDa [B].
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3475697&req=5

pone-0047789-g007: Antibody test by Western blot.Analyzed was A) lung tissue which is used as positive control for surfactant proteins; B) recombinantly synthesized SP-G protein (not purified) at 28°C (1) and 37°C (2), arrows indicate positive evidence of the surfactant protein G. The proteins extracted from the lung tissue and separated by 15% SDS-PAGE under reducing conditions show distinct bands for SP-G at the theoretically expected molecular weights of 11, 20 and 30 kDA [A]. In case of recombinantly expressed SP-G protein, the antibody detects a distinct band at 11 kDa [B].
Mentions: The specificity of the resulting antibody was tested with protein from lung tissue (30 µg) and with the recombinantly synthesized SP-G protein (not purified, 30 µg) (Figure 7). The purified antibody shows distinct protein bands in lung for SP-G at 11 kDa, 20 kDa and 30 kDa and a distinct protein band for recombinantly synthesized SP-G at about 12 kDa. We used lung tissue as specific positive control for surfactant proteins.

Bottom Line: In this work, computational chemistry and molecular-biological methods were combined to localize and characterize SP-G.With the help of a protein structure model, specific antibodies were obtained which allowed the detection of SP-G not only on mRNA but also on protein level.This includes also the possibility of interactions with lipid systems and with that, a potential surface-regulatory feature of SP-G.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioorganic Chemistry, Leibniz Institute of Plant Biochemistry, Halle, Germany.

ABSTRACT
Surfactant proteins (SP) are well known from human lung. These proteins assist the formation of a monolayer of surface-active phospholipids at the liquid-air interface of the alveolar lining, play a major role in lowering the surface tension of interfaces, and have functions in innate and adaptive immune defense. During recent years it became obvious that SPs are also part of other tissues and fluids such as tear fluid, gingiva, saliva, the nasolacrimal system, and kidney. Recently, a putative new surfactant protein (SFTA2 or SP-G) was identified, which has no sequence or structural identity to the already know surfactant proteins. In this work, computational chemistry and molecular-biological methods were combined to localize and characterize SP-G. With the help of a protein structure model, specific antibodies were obtained which allowed the detection of SP-G not only on mRNA but also on protein level. The localization of this protein in different human tissues, sequence based prediction tools for posttranslational modifications and molecular dynamic simulations reveal that SP-G has physicochemical properties similar to the already known surfactant proteins B and C. This includes also the possibility of interactions with lipid systems and with that, a potential surface-regulatory feature of SP-G. In conclusion, the results indicate SP-G as a new surfactant protein which represents an until now unknown surfactant protein class.

Show MeSH
Related in: MedlinePlus