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 journalArticlepeer-review

33 Scopus citations

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

All Science Journal Classification (ASJC) codes

  • Biochemistry

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