Linked Life Data

16551636

Source:http://linkedlifedata.com/resource/pubmed/id/16551636

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
22
pubmed:dateCreated
2006-5-29
pubmed:databankReference
pubmed:abstractText
Anti-DNA antibodies (Abs) are of biomedical interest because they are associated with autoimmune diseases in human and mice. Previously we isolated an anti-DNA monoclonal Ab 3D8 from an autoimmune-prone MRL-lpr/lpr mouse. Here we have characterized DNA binding kinetics and hydrolyzing activities of the recombinant single chain variable fragment (scFv) and the single variable domains of heavy chain (VH) and light chain (VL) using various single-stranded (ss) and double-stranded (ds) DNA substrates. All the Abs bound to both ds- and ssDNAs without significant preferential sequence specificity showing scFv higher affinities (KD = approximately 17-74 nm) than VH (KD = approximately 2.4-8.4 microm) and VL (KD = approximately 3.2-72 microm), and efficiently hydrolyzed both ds- and ssDNAs without sequence specificity in a Mg2+-dependent manner, except for the poor activity of 3D8 scFv for ss-(dT)40. Elucidated crystal structure-based His to Ala mutations on the complementarity determining regions of VH (His-H35 --> Ala) and/or VL (His-L94 --> Ala) of 3D8 scFv significantly inhibited the catalytic activities, indicating that the His residues are involved in the catalytic mechanism of 3D8 scFv. However, the DNA hydrolyzing activities of single domain VH and VL were not affected by the mutations, indicative of their different catalytic mechanisms from that of 3D8 scFv. Our results demonstrate single domain Abs with DNase activities for the first time, which might provide new insights into substrate recognition and catalytic mechanisms of anti-DNA Abs.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
2
pubmed:volume
281
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
15287-95
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:16551636-Amino Acid Substitution, pubmed-meshheading:16551636-Animals, pubmed-meshheading:16551636-Antibodies, Antinuclear, pubmed-meshheading:16551636-Base Sequence, pubmed-meshheading:16551636-Crystallography, X-Ray, pubmed-meshheading:16551636-DNA, pubmed-meshheading:16551636-DNA, Single-Stranded, pubmed-meshheading:16551636-Humans, pubmed-meshheading:16551636-Hydrolysis, pubmed-meshheading:16551636-Immunoglobulin Fragments, pubmed-meshheading:16551636-Immunoglobulin Heavy Chains, pubmed-meshheading:16551636-Immunoglobulin Light Chains, pubmed-meshheading:16551636-Kinetics, pubmed-meshheading:16551636-Mice, pubmed-meshheading:16551636-Mice, Inbred MRL lpr, pubmed-meshheading:16551636-Models, Molecular, pubmed-meshheading:16551636-Mutagenesis, Site-Directed, pubmed-meshheading:16551636-Protein Conformation, pubmed-meshheading:16551636-Protein Structure, Tertiary, pubmed-meshheading:16551636-Recombinant Proteins
pubmed:year
2006
pubmed:articleTitle
Heavy and light chain variable single domains of an anti-DNA binding antibody hydrolyze both double- and single-stranded DNAs without sequence specificity.
pubmed:affiliation
Department of Microbiology, Ajou University School of Medicine, San 5, Woncheon-dong, Yeongtong-gu, Suwon 443-749, Korea.
pubmed:publicationType
Journal Article, In Vitro, Research Support, Non-U.S. Gov't, Research Support, N.I.H., Extramural