Linked Life Data

11373276

Source:http://linkedlifedata.com/resource/pubmed/id/11373276

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
30
pubmed:dateCreated
2001-7-23
pubmed:abstractText
Mammalian phospholipase D (PLD) plays a key role in several signal transduction pathways and is involved in many diverse functions. To elucidate the complex molecular regulation of PLD, we investigated PLD-binding proteins obtained from rat brain extract. Here we report that a 43-kDa protein in the rat brain, beta-actin, acts as a major PLD2 direct-binding protein as revealed by peptide mass fingerprinting in combination with matrix-assisted laser desorption ionization/time-of-flight mass spectrometry. We also determined that the region between amino acids 613 and 723 of PLD2 is required for the direct binding of beta-actin, using bacterially expressed glutathione S-transferase fusion proteins of PLD2 fragments. Intriguingly, purified beta-actin potently inhibited both phosphatidylinositol-4,5-bisphosphate- and oleate-dependent PLD2 activities in a concentration-dependent manner (IC50 = 5 nm). In a previous paper, we reported that alpha-actinin inhibited PLD2 activity in an interaction-dependent and an ADP-ribosylation factor 1 (ARF1)-reversible manner (Park, J. B., Kim, J. H., Kim, Y., Ha, S. H., Kim, J. H., Yoo, J.-S., Du, G., Frohman, M. A., Suh, P.-G., and Ryu, S. H. (2000) J. Biol. Chem. 275, 21295-21301). In vitro binding analyses showed that beta-actin could displace alpha-actinin binding to PLD2, demonstrating independent interaction between cytoskeletal proteins and PLD2. Furthermore, ARF1 could steer the PLD2 activity in a positive direction regardless of the inhibitory effect of beta-actin on PLD2. We also observed that beta-actin regulates PLD1 and PLD2 with similar binding and inhibitory potencies. Immunocytochemical and co-immunoprecipitation studies demonstrated the in vivo interaction between the two PLD isozymes and actin in cells. Taken together, these results suggest that the regulation of PLD by cytoskeletal proteins, beta-actin and alpha-actinin, and ARF1 may play an important role in cytoskeleton-related PLD functions.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
27
pubmed:volume
276
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
28252-60
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:11373276-Actins, pubmed-meshheading:11373276-Animals, pubmed-meshheading:11373276-Baculoviridae, pubmed-meshheading:11373276-Brain, pubmed-meshheading:11373276-COS Cells, pubmed-meshheading:11373276-Cell Line, pubmed-meshheading:11373276-Cytoskeleton, pubmed-meshheading:11373276-DNA, Complementary, pubmed-meshheading:11373276-Dose-Response Relationship, Drug, pubmed-meshheading:11373276-Glutathione Transferase, pubmed-meshheading:11373276-Immunohistochemistry, pubmed-meshheading:11373276-Inhibitory Concentration 50, pubmed-meshheading:11373276-Insects, pubmed-meshheading:11373276-PC12 Cells, pubmed-meshheading:11373276-Phosphatidylinositol 4,5-Diphosphate, pubmed-meshheading:11373276-Phospholipase D, pubmed-meshheading:11373276-Precipitin Tests, pubmed-meshheading:11373276-Protein Binding, pubmed-meshheading:11373276-Rats, pubmed-meshheading:11373276-Recombinant Fusion Proteins, pubmed-meshheading:11373276-Spectrometry, Mass, Matrix-Assisted Laser..., pubmed-meshheading:11373276-Transfection
pubmed:year
2001
pubmed:articleTitle
Actin directly interacts with phospholipase D, inhibiting its activity.
pubmed:affiliation
Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang 790-784, Republic of Korea.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't