A distant evolutionary relationship between bacterial sphingomyelinase and mammalian DNase I

Yo Matsuo, Atsuko Yamada, Kikuo Tsukamoto, Hiroomi Tamura, Hiroh Ikezawa, Haruki Nakamura, Ken Nishikawa

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

The three-dimensional structure of bacterial sphingomyelinase (SMase) was predicted using a protein fold recognition method: the search of a library of known structures showed that the SMase sequence is highly compatible with the mammalian DNase 1 structure, which suggested that SMase adopts a structure similar to that of DNase I. The amino acid sequence alignment based on the prediction revealed that, despite the lack of overall sequence similarity (less than 10% identity), those residues of DNase I that are involved in the hydrolysis of the phosphodiester bond, including two histidine residues (His 134 and His 252) of the active center; are conserved in SMase. In addition, a conserved pentapeptide sequence motif was found, which includes two catalytically critical residues. Asp 251 and His 252. A sequence database search showed that the motif is highly specific to mammalian DNase I and bacterial SMase. The functional roles of SMase residues identified by the sequence comparison were consistent with the results from mutant studies. Two Bacillus cereus SMase mutants (H134A and H252A) were constructed by site-directed mutagenesis. They completely abolished their catalytic activity. A model for the SMase-sphingomyelin complex structure was built to investigate how the SMase specifically recognizes its substrate. The model suggested that a set of residues conserved among bacterial SMases, including Trp 28 and Phe 55, might be important in the substrate recognition. The predicted structural similarity and the conservation of the functionally important residues strongly suggest a distant evolutionary relationship between bacterial SMase and mammalian DNase I. These two phosphodiesterases must have acquired the specificity for different substrates in the course of evolution.

Original languageEnglish
Pages (from-to)2459-2467
Number of pages9
JournalProtein Science
Volume5
Issue number12
Publication statusPublished - 1996 Dec

Fingerprint

Sphingomyelin Phosphodiesterase
Deoxyribonuclease I
Substrates
Bacillus cereus
Bacterial Structures
Mutagenesis
Sphingomyelins
Deoxyribonucleases
Conserved Sequence
Sequence Alignment
Phosphoric Diester Hydrolases
Substrate Specificity
Site-Directed Mutagenesis
Histidine
Libraries
Amino Acid Sequence
Hydrolysis
Catalyst activity
Conservation
Databases

Keywords

  • bacterial sphingomyelinase
  • DNase I
  • evolutionary relationship
  • site- directed mutagenesis
  • structure prediction

ASJC Scopus subject areas

  • Biochemistry

Cite this

Matsuo, Y., Yamada, A., Tsukamoto, K., Tamura, H., Ikezawa, H., Nakamura, H., & Nishikawa, K. (1996). A distant evolutionary relationship between bacterial sphingomyelinase and mammalian DNase I. Protein Science, 5(12), 2459-2467.

A distant evolutionary relationship between bacterial sphingomyelinase and mammalian DNase I. / Matsuo, Yo; Yamada, Atsuko; Tsukamoto, Kikuo; Tamura, Hiroomi; Ikezawa, Hiroh; Nakamura, Haruki; Nishikawa, Ken.

In: Protein Science, Vol. 5, No. 12, 12.1996, p. 2459-2467.

Research output: Contribution to journalArticle

Matsuo, Y, Yamada, A, Tsukamoto, K, Tamura, H, Ikezawa, H, Nakamura, H & Nishikawa, K 1996, 'A distant evolutionary relationship between bacterial sphingomyelinase and mammalian DNase I', Protein Science, vol. 5, no. 12, pp. 2459-2467.
Matsuo Y, Yamada A, Tsukamoto K, Tamura H, Ikezawa H, Nakamura H et al. A distant evolutionary relationship between bacterial sphingomyelinase and mammalian DNase I. Protein Science. 1996 Dec;5(12):2459-2467.
Matsuo, Yo ; Yamada, Atsuko ; Tsukamoto, Kikuo ; Tamura, Hiroomi ; Ikezawa, Hiroh ; Nakamura, Haruki ; Nishikawa, Ken. / A distant evolutionary relationship between bacterial sphingomyelinase and mammalian DNase I. In: Protein Science. 1996 ; Vol. 5, No. 12. pp. 2459-2467.
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