Domain III of calpain is a Ca2+-regulated phospholipid-binding domain

Peter Tompa, Yasufumi Emori, Hiroyuki Sorimachi, Koichi Suzuki, P. Friedrich

Research output: Contribution to journalArticle

128 Citations (Scopus)

Abstract

The X-ray structure of m-calpain shows that domain III of the large subunit is structurally related to C2 domains, Ca2+-regulated lipid binding modules in many enzymes. To address whether this structural similarity entails functional analogy, we have characterized recombinant domain III from rat μ-and m-calpain and Drosophila CALPB. In a Ca2+ overlay assay domain III displays a large capacity for Ca2+ binding, commensurable with that of domain IV, the principal Ca2+-binding domain of calpains. The amount of Ca2+ bound to domain III increases 2- to 10-fold upon the addition of liposomes containing 20-40% di- and triphosphoinositides. Conversely, phospholipid-binding in spin-column size-exclusion chromatography is significantly promoted by Ca2+, in a manner similar to known C2 domains. These results suggest that domain III might be the primary lipid binding site of calpain and may play a decisive role in orchestrating Ca2+- and lipid activation of the enzyme.

Original languageEnglish
Pages (from-to)1333-1339
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume280
Issue number5
DOIs
Publication statusPublished - 2001

Fingerprint

Calpain
Phospholipids
Lipids
Column chromatography
Enzyme Activation
Size exclusion chromatography
Enzymes
Liposomes
Drosophila
Gel Chromatography
Rats
Assays
Chemical activation
Binding Sites
X-Rays
X rays
m-calpain
C2 Domains

Keywords

  • C2 domain
  • Ca binding
  • Calpain
  • Domain III
  • Phospholipid binding

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Domain III of calpain is a Ca2+-regulated phospholipid-binding domain. / Tompa, Peter; Emori, Yasufumi; Sorimachi, Hiroyuki; Suzuki, Koichi; Friedrich, P.

In: Biochemical and Biophysical Research Communications, Vol. 280, No. 5, 2001, p. 1333-1339.

Research output: Contribution to journalArticle

Tompa, Peter ; Emori, Yasufumi ; Sorimachi, Hiroyuki ; Suzuki, Koichi ; Friedrich, P. / Domain III of calpain is a Ca2+-regulated phospholipid-binding domain. In: Biochemical and Biophysical Research Communications. 2001 ; Vol. 280, No. 5. pp. 1333-1339.
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