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dc.contributor.author Hard, RL
dc.contributor.author Liu, J
dc.contributor.author Shen, J
dc.contributor.author Zhou, P
dc.contributor.author Pei, D
dc.coverage.spatial United States
dc.date.accessioned 2011-06-21T17:22:10Z
dc.date.issued 2010-12-21
dc.identifier http://www.ncbi.nlm.nih.gov/pubmed/21090589
dc.identifier.citation Biochemistry, 2010, 49 (50), pp. 10737 - 10746
dc.identifier.uri http://hdl.handle.net/10161/4013
dc.description.abstract The BUZ/Znf-UBP domain is a protein module found in the cytoplasmic deacetylase HDAC6, E3 ubiquitin ligase BRAP2/IMP, and a subfamily of ubiquitin-specific proteases. Although several BUZ domains have been shown to bind ubiquitin with high affinity by recognizing its C-terminal sequence (RLRGG-COOH), it is currently unknown whether the interaction is sequence-specific or whether the BUZ domains are capable of binding to proteins other than ubiquitin. In this work, the BUZ domains of HDAC6 and Ubp-M were subjected to screening against a one-bead-one-compound (OBOC) peptide library that exhibited random peptide sequences with free C-termini. Sequence analysis of the selected binding peptides as well as alanine scanning studies revealed that the BUZ domains require a C-terminal Gly-Gly motif for binding. At the more N-terminal positions, the two BUZ domains have distinct sequence specificities, allowing them to bind to different peptides and/or proteins. A database search of the human proteome on the basis of the BUZ domain specificities identified 11 and 24 potential partner proteins for Ubp-M and HDAC6 BUZ domains, respectively. Peptides corresponding to the C-terminal sequences of four of the predicted binding partners (FBXO11, histone H4, PTOV1, and FAT10) were synthesized and tested for binding to the BUZ domains by fluorescence polarization. All four peptides bound to the HDAC6 BUZ domain with low micromolar K(D) values and less tightly to the Ubp-M BUZ domain. Finally, in vitro pull-down assays showed that the Ubp-M BUZ domain was capable of binding to the histone H3-histone H4 tetramer protein complex. Our results suggest that BUZ domains are sequence-specific protein-binding modules, with each BUZ domain potentially binding to a different subset of proteins.
dc.format.extent 10737 - 10746
dc.language ENG
dc.language.iso en_US en_US
dc.relation.ispartof Biochemistry
dc.relation.isversionof 10.1021/bi101014s
dc.subject Amino Acid Sequence
dc.subject Histone Deacetylases
dc.subject Humans
dc.subject Peptide Library
dc.subject Protein Structure, Tertiary
dc.subject Proteins
dc.subject Ubiquitin Thiolesterase
dc.title HDAC6 and Ubp-M BUZ domains recognize specific C-terminal sequences of proteins.
dc.title.alternative en_US
dc.type Journal Article
dc.description.version Version of Record en_US
duke.date.pubdate 2010-12-21 en_US
duke.description.endpage 10746 en_US
duke.description.issue 50 en_US
duke.description.startpage 10737 en_US
duke.description.volume 49 en_US
dc.relation.journal Biochemistry en_US
pubs.author-url http://www.ncbi.nlm.nih.gov/pubmed/21090589
pubs.issue 50
pubs.organisational-group /Duke
pubs.organisational-group /Duke/School of Medicine
pubs.organisational-group /Duke/School of Medicine/Basic Science Departments
pubs.organisational-group /Duke/School of Medicine/Basic Science Departments/Biochemistry
pubs.organisational-group /Duke/School of Medicine/Institutes and Centers
pubs.organisational-group /Duke/School of Medicine/Institutes and Centers/Duke Cancer Institute
pubs.organisational-group /Duke/Trinity College of Arts & Sciences
pubs.organisational-group /Duke/Trinity College of Arts & Sciences/Chemistry
pubs.publication-status Published
pubs.volume 49
dc.identifier.eissn 1520-4995

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