Linked Life Data

9593187

Source:http://linkedlifedata.com/resource/pubmed/id/9593187

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1998-7-1
pubmed:abstractText
A recombinant Fv construct of the B1 monoclonal antibody that recognizes the LewisY-related carbohydrate epitope on human carcinoma cells has been prepared. The Fv is composed of the polypeptide chains of the VH and VL domains expressed independently and isolated as inclusion bodies. The Fv is prepared by combining and refolding equimolar amounts of guanidine chloride solubilized inclusion bodies. The Fv is stabilized by an engineered interchain disulfide bridge between residues VL100 and VH44. This construct has a similar binding affinity as that of the single-chain construct (Benhar and Pastan, Clin. Cancer Res. 1:1023-1029, 1995). The B1 disulfide-stabilized Fv (BldsFv) crystallizes in space group P6(1)22 with the unit cell parameters a = b = 80.1 A, and c = 138.1 A. The crystal structure of the BldsFv has been determined at 2.1-A resolution using the molecular replacement technique. The final structure has a crystallographic R-value of 0.187 with a root mean square deviation in bond distance of 0.014 A and in bond angle of 2.74 degrees. Comparisons of the BldsFv structure with known structures of Fv regions of other immunoglobulin fragments shows closely related secondary and tertiary structures. The antigen combining site of BldsFv is a deep depression 10-A wide and 17-A long with the walls of the depression composed of residues, many of which are tyrosines, from complementarity determining regions L1, L3, H1, H2, and H3. Model building studies indicate that the LewisY tetrasaccharide, Fuc-Gal-Nag-Fuc, can be accommodated in the antigen combining site in a manner consistent with the epitope predicted in earlier biochemical studies (Pastan, Lovelace, Gallo, Rutherford, Magnani, and Willingham, Cancer Res. 51:3781-3787, 1991). Thus, the engineered disulfide bridge appears to cause little, if any, distortion in the Fv structure, making it an effective substitute for the B1 Fab.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
0887-3585
pubmed:author
pubmed:issnType
Print
pubmed:day
1
pubmed:volume
31
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
128-38
pubmed:dateRevised
2011-11-17
pubmed:meshHeading
pubmed-meshheading:9593187-Amino Acid Sequence, pubmed-meshheading:9593187-Animals, pubmed-meshheading:9593187-Antibodies, Monoclonal, pubmed-meshheading:9593187-Antibodies, Neoplasm, pubmed-meshheading:9593187-Antigen-Antibody Reactions, pubmed-meshheading:9593187-Binding Sites, Antibody, pubmed-meshheading:9593187-Crystallography, X-Ray, pubmed-meshheading:9593187-Cystine, pubmed-meshheading:9593187-Epitopes, pubmed-meshheading:9593187-Immunoglobulin Fragments, pubmed-meshheading:9593187-Immunoglobulin G, pubmed-meshheading:9593187-Lewis Blood-Group System, pubmed-meshheading:9593187-Mice, pubmed-meshheading:9593187-Models, Molecular, pubmed-meshheading:9593187-Molecular Sequence Data, pubmed-meshheading:9593187-Protein Conformation, pubmed-meshheading:9593187-Protein Structure, Secondary, pubmed-meshheading:9593187-Protein Structure, Tertiary
pubmed:year
1998
pubmed:articleTitle
Crystal structure of the disulfide-stabilized Fv fragment of anticancer antibody B1: conformational influence of an engineered disulfide bond.
pubmed:affiliation
Center for Advanced Research in Biotechnology, University of Maryland Biotechnology Institute and the National Institute of Standards and Technology, Rockville, Maryland 20850, USA.
pubmed:publicationType
Journal Article