Effect of cysteine 85 on biochemical properties and biological function of human surfactant protein A variants

Guirong Wang, Catherine Myers, Anatoly Mikerov, Joanna Floros

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Four "core" amino acid differences within the collagen-like domain distinguish the human surfactant protein A1 (SP-A1) variants from the SP-A2 variants. One of these, cysteine 85 that could form intermolecular disulfide bonds, is present in SP-A1 (Cys85) and absent in SP-A2 (Arg85). We hypothesized that residue 85 affects both the structure and function of SP-A1 and SP-A2 variants. To test this, wild-type (WT) variants, 6A2 of SP-A1 and 1A0 of SP-A2, and their mutants (6A 2(C85R) and 1A0(R85C)) were generated and studied. We found the following: (1) Residue 85 affected the binding ability to mannose and the oligomerization pattern of SP-As. The 1A0(R85C) and 6A 2(C85R) patterns were similar and/or resembled those of WT 6A 2 and 1A0, respectively. (2) Both SP-A WT and mutants differentially induced rough LPS and Pseudomonas aeruginosa aggregation in the following order: 1A0 > 6A2 > 6A2(C85R) > 1A0(R85C) for Re-LPS aggregation and 1A0 > 6A 2 = 6A2(C85R) = 1A0(R85C) for bacterial aggregation. (3) SP-A WT and mutants enhanced phagocytosis of P. aeruginosa by rat alveolar macrophages. Their phagocytic index order was 6A2(C85R) > 1A0 > 6A2 = 1A0(R85C). The activity of mutant 1A0(C85R) was significantly lower than WT 1A0 but similar to 6A2. Compared to WT 6A2, the 6A 2(C85R) mutant exhibited a significantly higher activity. These results indicate that the SP-A variant/mutant with Arg85 exhibits a higher ability to enhance bacterial phagocytosis than that with Cys 85. Residue 85 plays an important role in the structure and function of SP-A and is a major factor for the differences between SP-A1 and SP-A2 variants.

Original languageEnglish (US)
Pages (from-to)8425-8435
Number of pages11
JournalBiochemistry
Volume46
Issue number28
DOIs
StatePublished - Jul 17 2007

Fingerprint

Pulmonary Surfactant-Associated Protein A
varespladib methyl
Surface-Active Agents
Cysteine
Aptitude
Agglomeration
Proteins
Phagocytosis
Pseudomonas aeruginosa
Oligomerization
Alveolar Macrophages
Mannose
Disulfides
Rats
Collagen
Amino Acids

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Wang, Guirong ; Myers, Catherine ; Mikerov, Anatoly ; Floros, Joanna. / Effect of cysteine 85 on biochemical properties and biological function of human surfactant protein A variants. In: Biochemistry. 2007 ; Vol. 46, No. 28. pp. 8425-8435.
@article{8ad20ee08dca4bc5951d895742adfad2,
title = "Effect of cysteine 85 on biochemical properties and biological function of human surfactant protein A variants",
abstract = "Four {"}core{"} amino acid differences within the collagen-like domain distinguish the human surfactant protein A1 (SP-A1) variants from the SP-A2 variants. One of these, cysteine 85 that could form intermolecular disulfide bonds, is present in SP-A1 (Cys85) and absent in SP-A2 (Arg85). We hypothesized that residue 85 affects both the structure and function of SP-A1 and SP-A2 variants. To test this, wild-type (WT) variants, 6A2 of SP-A1 and 1A0 of SP-A2, and their mutants (6A 2(C85R) and 1A0(R85C)) were generated and studied. We found the following: (1) Residue 85 affected the binding ability to mannose and the oligomerization pattern of SP-As. The 1A0(R85C) and 6A 2(C85R) patterns were similar and/or resembled those of WT 6A 2 and 1A0, respectively. (2) Both SP-A WT and mutants differentially induced rough LPS and Pseudomonas aeruginosa aggregation in the following order: 1A0 > 6A2 > 6A2(C85R) > 1A0(R85C) for Re-LPS aggregation and 1A0 > 6A 2 = 6A2(C85R) = 1A0(R85C) for bacterial aggregation. (3) SP-A WT and mutants enhanced phagocytosis of P. aeruginosa by rat alveolar macrophages. Their phagocytic index order was 6A2(C85R) > 1A0 > 6A2 = 1A0(R85C). The activity of mutant 1A0(C85R) was significantly lower than WT 1A0 but similar to 6A2. Compared to WT 6A2, the 6A 2(C85R) mutant exhibited a significantly higher activity. These results indicate that the SP-A variant/mutant with Arg85 exhibits a higher ability to enhance bacterial phagocytosis than that with Cys 85. Residue 85 plays an important role in the structure and function of SP-A and is a major factor for the differences between SP-A1 and SP-A2 variants.",
author = "Guirong Wang and Catherine Myers and Anatoly Mikerov and Joanna Floros",
year = "2007",
month = "7",
day = "17",
doi = "10.1021/bi7004569",
language = "English (US)",
volume = "46",
pages = "8425--8435",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "28",

}

Effect of cysteine 85 on biochemical properties and biological function of human surfactant protein A variants. / Wang, Guirong; Myers, Catherine; Mikerov, Anatoly; Floros, Joanna.

In: Biochemistry, Vol. 46, No. 28, 17.07.2007, p. 8425-8435.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Effect of cysteine 85 on biochemical properties and biological function of human surfactant protein A variants

AU - Wang, Guirong

AU - Myers, Catherine

AU - Mikerov, Anatoly

AU - Floros, Joanna

PY - 2007/7/17

Y1 - 2007/7/17

N2 - Four "core" amino acid differences within the collagen-like domain distinguish the human surfactant protein A1 (SP-A1) variants from the SP-A2 variants. One of these, cysteine 85 that could form intermolecular disulfide bonds, is present in SP-A1 (Cys85) and absent in SP-A2 (Arg85). We hypothesized that residue 85 affects both the structure and function of SP-A1 and SP-A2 variants. To test this, wild-type (WT) variants, 6A2 of SP-A1 and 1A0 of SP-A2, and their mutants (6A 2(C85R) and 1A0(R85C)) were generated and studied. We found the following: (1) Residue 85 affected the binding ability to mannose and the oligomerization pattern of SP-As. The 1A0(R85C) and 6A 2(C85R) patterns were similar and/or resembled those of WT 6A 2 and 1A0, respectively. (2) Both SP-A WT and mutants differentially induced rough LPS and Pseudomonas aeruginosa aggregation in the following order: 1A0 > 6A2 > 6A2(C85R) > 1A0(R85C) for Re-LPS aggregation and 1A0 > 6A 2 = 6A2(C85R) = 1A0(R85C) for bacterial aggregation. (3) SP-A WT and mutants enhanced phagocytosis of P. aeruginosa by rat alveolar macrophages. Their phagocytic index order was 6A2(C85R) > 1A0 > 6A2 = 1A0(R85C). The activity of mutant 1A0(C85R) was significantly lower than WT 1A0 but similar to 6A2. Compared to WT 6A2, the 6A 2(C85R) mutant exhibited a significantly higher activity. These results indicate that the SP-A variant/mutant with Arg85 exhibits a higher ability to enhance bacterial phagocytosis than that with Cys 85. Residue 85 plays an important role in the structure and function of SP-A and is a major factor for the differences between SP-A1 and SP-A2 variants.

AB - Four "core" amino acid differences within the collagen-like domain distinguish the human surfactant protein A1 (SP-A1) variants from the SP-A2 variants. One of these, cysteine 85 that could form intermolecular disulfide bonds, is present in SP-A1 (Cys85) and absent in SP-A2 (Arg85). We hypothesized that residue 85 affects both the structure and function of SP-A1 and SP-A2 variants. To test this, wild-type (WT) variants, 6A2 of SP-A1 and 1A0 of SP-A2, and their mutants (6A 2(C85R) and 1A0(R85C)) were generated and studied. We found the following: (1) Residue 85 affected the binding ability to mannose and the oligomerization pattern of SP-As. The 1A0(R85C) and 6A 2(C85R) patterns were similar and/or resembled those of WT 6A 2 and 1A0, respectively. (2) Both SP-A WT and mutants differentially induced rough LPS and Pseudomonas aeruginosa aggregation in the following order: 1A0 > 6A2 > 6A2(C85R) > 1A0(R85C) for Re-LPS aggregation and 1A0 > 6A 2 = 6A2(C85R) = 1A0(R85C) for bacterial aggregation. (3) SP-A WT and mutants enhanced phagocytosis of P. aeruginosa by rat alveolar macrophages. Their phagocytic index order was 6A2(C85R) > 1A0 > 6A2 = 1A0(R85C). The activity of mutant 1A0(C85R) was significantly lower than WT 1A0 but similar to 6A2. Compared to WT 6A2, the 6A 2(C85R) mutant exhibited a significantly higher activity. These results indicate that the SP-A variant/mutant with Arg85 exhibits a higher ability to enhance bacterial phagocytosis than that with Cys 85. Residue 85 plays an important role in the structure and function of SP-A and is a major factor for the differences between SP-A1 and SP-A2 variants.

UR - http://www.scopus.com/inward/record.url?scp=34447561643&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34447561643&partnerID=8YFLogxK

U2 - 10.1021/bi7004569

DO - 10.1021/bi7004569

M3 - Article

VL - 46

SP - 8425

EP - 8435

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 28

ER -