Drosophila UNC-45 prevents heat-induced aggregation of skeletal muscle myosin and facilitates refolding of citrate synthase

Girish C. Melkani, Chi F. Lee, Anthony Ross Cammarato, Sanford I. Bernstein

Research output: Contribution to journalArticle

Abstract

UNC-45 belongs to the UCS (UNC-45, CRO1, She4p) domain protein family, whose members interact with various classes of myosin. Here we provide structural and biochemical evidence that Escherichia coli-expressed Drosophila UNC-45 (DUNC-45) maintains the integrity of several substrates during heat-induced stress in vitro. DUNC-45 displays chaperone function in suppressing aggregation of the muscle myosin heavy meromyosin fragment, the myosin S-1 motor domain, α-lactalbumin and citrate synthase. Biochemical evidence is supported by electron microscopy, which reveals the first structural evidence that DUNC-45 prevents inter- or intra-molecular aggregates of skeletal muscle heavy meromyosin caused by elevated temperatures. We also demonstrate for the first time that UNC-45 is able to refold a denatured substrate, urea-unfolded citrate synthase. Overall, this in vitro study provides insight into the fate of muscle myosin under stress conditions and suggests that UNC-45 protects and maintains the contractile machinery during in vivo stress.

Original languageEnglish (US)
Pages (from-to)317-322
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume396
Issue number2
DOIs
StatePublished - May 28 2010
Externally publishedYes

Fingerprint

Skeletal Muscle Myosins
Myosin Subfragments
Citrate (si)-Synthase
Myosins
Drosophila
Muscle
Agglomeration
Hot Temperature
Lactalbumin
Muscles
Substrates
Escherichia coli
Electron microscopy
Machinery
Urea
Electron Microscopy
Skeletal Muscle
Temperature
Proteins
In Vitro Techniques

Keywords

  • Chaperone
  • Drosophila
  • Myosin
  • Protein aggregation
  • Refolding
  • UNC-45

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Drosophila UNC-45 prevents heat-induced aggregation of skeletal muscle myosin and facilitates refolding of citrate synthase. / Melkani, Girish C.; Lee, Chi F.; Cammarato, Anthony Ross; Bernstein, Sanford I.

In: Biochemical and Biophysical Research Communications, Vol. 396, No. 2, 28.05.2010, p. 317-322.

Research output: Contribution to journalArticle

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