Structural Analysis and Lipid-Binding Properties of Recombinant Human Surfactant Protein A Derived from One or Both Genes

García-Verdugo, Ignacio; Wang, G.; Floros, Joanna; Casals, Cristina

doi:10.1021/bi026540l

Cited by 66 publications

(156 citation statements)

References 60 publications

Supporting

Mentioning

141

Contrasting

Order By: Relevance

“…Studies of host/guest triple helical peptides, where a single guest Gly-X-Y with substitutions in X and Y positions was studied while embedded in a collagen host peptide, showed that an Arg in the Y position associates with high stability of the host/guest peptide (9,61,73). Based on these studies, we have previously proposed (21,22) that Arg 85 (SP-A2) has a higher local microstability compared with Cys 85 (SP-A1). This may result in local "microunfolding" and exposure of SP-A1- specific epitopes.…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, qualitative and quantitative differences have been observed in vitro between the SP-A1 and SP-A2 genes (22,36,57,60,(77)(78)(79)(80)(81), pointing to the possibility that the overall functional activity of SP-A may depend on the relative SP-A1-to-SP-A2 levels rather than on the total SP-A content. To address this possibility, it .…”

Section: Discussionmentioning

confidence: 99%

“…The primary structure of SP-A in the collagenous domain consists of 23 Gly-X-Y repeats, where Gly represents glycine and X or Y represents any other aa residue. However, an important difference exists between SP-A1 and SP-A2 in Gly-X-Y repeat number 18 (16,22). SP-A1 has a Cys in the Y position (GEC) referred to as Cys 85 , whereas the SP-A2 has an Arg (GER) referred to as Arg 85 .…”

Section: Discussionmentioning

confidence: 99%

“…More than 30 alleles have been characterized for these two genes, with four SP-A1 alleles (6A, 6A 2 , 6A 3 , and 6A 4 ) and six SP-A2 alleles (1A, 1A 0 , 1A 1 , 1A 2 , 1A 3 , and 1A 5 ) most frequently observed in the general population (10,14,16,40,67). Although a high level of sequence similarity is shared among the SP-A genes and their corresponding alleles, in vitro studies have identified both qualitative (22,36,57,60,77,80,81) and quantitative (24,35,38,46,56,69,70,78,79) differences between the two genes.…”

mentioning

confidence: 99%

“…Functionally, SP-A2 exhibits a higher level of activity than SP-A1 in its ability to enhance TNF-␣ and IL-8 production by the macrophage-like THP-1 cell line (80,81), phagocytosis of Pseudomonas aeruginosa by rat alveolar macrophages (57), and secretion of phosphatidylcholine by type II alveolar cells (77). Structurally, circular dichroic spectroscopy studies have shown SP-A2 to be more stable than SP-A1 (22). Biochemical differences in response to ozone exposure (36,77,81), oligomerization patterns (36,77), aggregation (71), and the ability to bind carbohydrates (60) were also observed for SP-A1 and SP-A2 variants.…”

mentioning

confidence: 99%

See 4 more Smart Citations

Characterization of a human surfactant protein A1 (SP-A1) gene-specific antibody; SP-A1 content variation among individuals of varying age and pulmonary health

Tagaram¹,

Wang²,

Umstead³

et al. 2007

American Journal of Physiology-Lung Cellular and Molecular Phys

View full text Add to dashboard Cite

The human surfactant protein A (SP-A) locus consists of two functional genes (SP-A1, SP-A2) with gene-specific products exhibiting qualitative and quantitative differences. The aim here was twofold: 1) generate SP-A1 genespecific antibody, and 2) use this to assess gene-specific SP-A content in the bronchoalveolar lavage fluid (BALF). An SP-A1-specific polyclonal antibody (hSP-A1_Ab 68-88 _Col) was raised in chicken, and its specificity was determined by immunoblot and ELISA using mammalian Chinese hamster ovary (CHO) cell-expressed SP-A1 and SP-A2 variants and by immunofluorescence with stably transfected CHO cell lines expressing SP-A1 or SP-A2 variants. SP-A1 content was evaluated according to age and lung status. A gradual decrease (P Ͻ 0.05) in SP-A1/SP-A ratio was observed in healthy subjects (HS) with increased age, although no significant change was observed in total SP-A content among age groups. Total SP-A and SP-A1 content differed significantly between alveolar proteinosis (AP) patients and HS, with no significant difference observed in SP-A1/SP-A ratio between AP and HS. The cystic fibrosis (CF) ratio was significantly higher compared with AP, HS, and noncystic fibrosis (NCF), even though SP-A1 and total SP-A were decreased in CF compared with most of the other groups. The ratio was higher in culture-positive vs. culture-negative samples from CF and NCF (P ϭ 0.031). A trend of an increased ratio was observed in culture-positive CF (0.590 Ϯ 0.10) compared with culture-positive NCF (0.368 Ϯ 0.085). In summary, we developed and characterized an SP-A1 gene-specific antibody and used it to identify gene-specific SP-A content in BALFs as a function of age and lung health. alveolar proteinosis; bronchoalveolar lavage fluid; cystic fibrosis; peptide antibody; surfactant protein A HUMAN SURFACTANT PROTEIN A (SP-A), a member of the C-type lectin or collectin family, is encoded by two functional genes, SP-A1 and SP-A2, with a high percentage of sequence similarity detected at the nucleotide (94%) and the amino acid (aa) (96%) levels. More than 30 alleles have been characterized for these two genes, with four SP-A1 alleles (6A, 6A 2 , 6A 3 , and 6A 4 ) and six SP-A2 alleles (1A, 1A 0 , 1A 1 , 1A 2 , 1A 3 , and 1A 5 ) most frequently observed in the general population (10,14,16,40,67). Although a high level of sequence similarity is shared among the SP-A genes and their corresponding alleles, in vitro studies have identified both qualitative (22,36,57,60,77,80,81) and quantitative (24,35,38,46,56,69,70,78,79) differences between the two genes.Qualitative differences between the SP-A1 and SP-A2 genes include functional, structural, and biochemical differences. Functionally, SP-A2 exhibits a higher level of activity than SP-A1 in its ability to enhance TNF-␣ and IL-8 production by the macrophage-like THP-1 cell line (80, 81), phagocytosis of Pseudomonas aeruginosa by rat alveolar macrophages (57), and secretion of phosphatidylcholine by type II alveolar cells (77). Structurally, circular dichroic spectroscopy stud...

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Characterization of a human surfactant protein A1 (SP-A1) gene-specific antibody; SP-A1 content variation among individuals of varying age and pulmonary health

Tagaram¹,

Wang²,

Umstead³

et al. 2007

American Journal of Physiology-Lung Cellular and Molecular Phys

View full text Add to dashboard Cite

show abstract

Structure and Function of Pulmonary Surfactant Proteins

García‐Álvarez

Alonso

Pérez‐Gil

2019

Encyclopedia of Life Sciences

View full text Add to dashboard Cite

Pulmonary surfactant is a lipid/protein complex essential to keep the airspaces of mammalian lungs open. It forms surface‐active films at the respiratory air–liquid interface, reducing surface tension to a minimum and thus minimising the work of breathing. At the same time, surfactant establishes the first barrier against the entry of potential pathogens through the large surface that lungs expose to environment. Both the interfacial and the innate defence activities depend critically on the presence of highly specialised evolutionarily conserved surfactant‐associated proteins. The structure and molecular mechanisms of these proteins have been extensively characterised in the last decades, in studies that constitute the basis for current models on surfactant mechanisms at the alveolar spaces, in health and disease. This article summarises the current knowledge on the structure–function relationships defining the molecular action of pulmonary surfactant proteins and how they are being key actors to maintain operative breathing in the lungs. Key Concepts The presence of specific proteins is essential for pulmonary surfactant to play its crucial role in stabilising the respiratory air–liquid interface. Collectin proteins SP‐A and SP‐D oligomerise to form trimers and supratrimeric oligomers with multivalent binding capacities to sugars and lipids and participate in innate defence mechanisms, binding to the surface of pathogens and allergens to facilitate their clearance. Surfactant protein SP‐B is essential for life; its absence is associated with an irreversible respiratory failure at birth. SP‐B belongs to the saposin protein family and assembles as ring‐shaped oligomers that enclose hydrophobic channels competent to transfer phospholipids between membranes and between membranes and the interface, avoiding its exposure to aqueous environments. Some synthetic peptides designed to mimic key SP‐B motifs are being used to produce alternative surfactant preparations for therapeutic applications. Protein SP‐C is a very hydrophobic helical protein whose expression is linked to the differentiation of lung tissue. Its oligomerisation is associated with surfactant membrane fragmentation and their interconversions at the alveolar spaces.

show abstract

Pulmonary SP-A: Forms and Functions

Gupta

2012

Animal Lectins: Form, Function and Clinical Applications

View full text Add to dashboard Cite

Structural Analysis and Lipid-Binding Properties of Recombinant Human Surfactant Protein A Derived from One or Both Genes

Cited by 66 publications

References 60 publications

Characterization of a human surfactant protein A1 (SP-A1) gene-specific antibody; SP-A1 content variation among individuals of varying age and pulmonary health

Characterization of a human surfactant protein A1 (SP-A1) gene-specific antibody; SP-A1 content variation among individuals of varying age and pulmonary health

Structure and Function of Pulmonary Surfactant Proteins

Pulmonary SP-A: Forms and Functions

Contact Info

Product

Resources

About