Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
8
pubmed:dateCreated
2003-7-22
pubmed:abstractText
Recently, it was shown that Yersinia outer protein T (YopT) belongs to a new family of cysteine proteases containing invariant C, H, and D residues that are crucial for its activity. YopT cleaves RhoA, Rac, and Cdc42 at their C termini, thereby releasing them from the membrane. Moreover, YopT inhibits the Rho-rhotekin and Rho-guanine nucleotide dissociation inhibitor interactions. To characterize the active domain of YopT, we constructed N- and C-terminal truncations and expressed them as glutathione S-transferase fusion proteins in Escherichia coli. The toxin fragments were tested for stability by trypsin digestion. The activity of the proteins was studied by membrane release assay, rhotekin pulldown experiments, and microinjection. Whereas deletion of the first 74 N-terminal amino acids did not influence the activity of YopT, deletion of 8 amino acids from the C terminus led to complete loss of activity. N-terminal deletion of 100 amino acids led to an inactive protein, although it still contained the amino acids C139, H258, and D274, which are essential for catalysis. Loss of activity of the N-terminal deletions corresponded to the block of interaction with RhoA, indicating that residues 75 to 100 of YopT are essential for binding to the GTPase. By contrast, when up to 15 amino acids of the C terminus were deleted, the protein had no activity but was still able to interact with RhoA, suggesting a role for the C terminus in the enzyme activity of YopT.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-10506187, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-10816416, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-10844661, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-10874740, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-11018537, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-11152757, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-11705930, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-11952637, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-12062101, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-12538863, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-8294016, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-9171345, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-9188492, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-9192900, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-9438836, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-9535085, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-9723929, http://linkedlifedata.com/resource/pubmed/commentcorrection/12874342-9841674
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0019-9567
pubmed:author
pubmed:issnType
Print
pubmed:volume
71
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
4623-32
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed-meshheading:12874342-Animals, pubmed-meshheading:12874342-Bacterial Proteins, pubmed-meshheading:12874342-Base Sequence, pubmed-meshheading:12874342-Carrier Proteins, pubmed-meshheading:12874342-Catalytic Domain, pubmed-meshheading:12874342-Cattle, pubmed-meshheading:12874342-Cells, Cultured, pubmed-meshheading:12874342-Cloning, Molecular, pubmed-meshheading:12874342-Cysteine Endopeptidases, pubmed-meshheading:12874342-Cytotoxins, pubmed-meshheading:12874342-DNA, Bacterial, pubmed-meshheading:12874342-Enzyme Stability, pubmed-meshheading:12874342-HeLa Cells, pubmed-meshheading:12874342-Humans, pubmed-meshheading:12874342-Intracellular Signaling Peptides and Proteins, pubmed-meshheading:12874342-Rats, pubmed-meshheading:12874342-Recombinant Fusion Proteins, pubmed-meshheading:12874342-Sequence Deletion, pubmed-meshheading:12874342-Yersinia, pubmed-meshheading:12874342-rhoA GTP-Binding Protein
pubmed:year
2003
pubmed:articleTitle
The C terminus of YopT is crucial for activity and the N terminus is crucial for substrate binding.
pubmed:affiliation
Institut für Experimentelle und Klinische Pharmakologie und Toxikologie, Albert-Ludwigs-Universität Freiburg, D-79104 Freiburg, Germany.
pubmed:publicationType
Journal Article, In Vitro, Research Support, Non-U.S. Gov't