Erythrocyte spectrin's chimeric E2/E3 ubiquitin conjugating/ligating activity

Y. J. Hsu, W. E. Zimmer, Steven Goodman

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Spectrin is important for the shape and the physical properties of the red blood cell, such as deformability and resistance to mechanical stress. Previous findings from our laboratory indicated that human erythrocyte alpha-spectrin can facilitate formation of ubiquitin-spectrin adducts and conjugates. Computer analysis revealed domains that contained significant homologies to known consensus catalytic E2 and E3 sequences, and allowed us to develop a model for alpha-spectrin ubiquitin conjugating enzyme (E2) and ubiquitin protein ligase (E3) enzymatic activities. In order to identify the precise E2/E3 site(s), in the present study, a GST-fusion alpha-spectrin (2005-2415) recombinant protein was tested using a cell free in vitro ubiquitination assay. We found that cysteine 2071 and cysteine 2100 are critical for alpha-spectrin (2005-2415) E2/E3 activity. Furthermore, together with testing an additional 13 site-specific mutants, we also demonstrated that both Cys2071 and Cys2100 are capable of transferring ubiquitin from an E1 enzyme to target sites within alpha-spectrin (2005-2415).

Original languageEnglish (US)
Pages (from-to)187-193
Number of pages7
JournalCellular and Molecular Biology
Volume51
Issue number2
StatePublished - Sep 5 2005
Externally publishedYes

Fingerprint

Spectrin
Ubiquitin
Erythrocytes
Cysteine
Ubiquitin-Conjugating Enzymes
Mechanical Stress
Ubiquitin-Protein Ligases
Ubiquitination
Formability
Recombinant Proteins
Assays
Blood
Fusion reactions
Physical properties
Cells
Testing
Enzymes

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Erythrocyte spectrin's chimeric E2/E3 ubiquitin conjugating/ligating activity. / Hsu, Y. J.; Zimmer, W. E.; Goodman, Steven.

In: Cellular and Molecular Biology, Vol. 51, No. 2, 05.09.2005, p. 187-193.

Research output: Contribution to journalArticle

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