Linked Life Data

2477068

Source:http://linkedlifedata.com/resource/pubmed/id/2477068

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1989-11-15
pubmed:abstractText
To understand the mechanism by which colipase acts as a protein cofactor for anchoring pancreatic lipase at triacylglycerol/water interface, we have used an immunochemical approach. Ten monoclonal antibodies (Mabs) against porcine pancreatic procolipase were produced. Purified immunoglobulins and Fab fragments were studied for their capacity to inhibit colipase-dependent lipase activity. These studies were carried out by using procolipase, the secretory form of the cofactor, and its trypsin-treated form obtained by removal of the amino terminal pentapeptide by trypsin. Reactivities of Mabs with both forms of the cofactor were also studied by immunoenzymatic methods. Mabs 6.1, 49.20. 75.8, 270.13 and 419.1 were found to inhibit lipolysis by preventing the binding of procolipase or trypsin-treated colipase to the lipid substrate. Mab 72.11 inhibited procolipase binding but had no effect on trypsin-treated colipase. Mab 72.11 reacted with procolipase in ELISA but showed no reactivity with trypsin-treated colipase. Finally, preincubation of Mab 72.11 with porcine procolipase prevented specific cleavage at the Arg5-Gly6 bond by trypsin. It could be concluded, that the five first residues of procolipase are structural elements of the antigenic determinant recognized by Mab 72.11. Results of ELISA additivity tests (cotitrations) further indicated that epitopes for Mabs 6.1, 72.11, 270.13 and 419.1 and for Mabs 49.20 and 75.8 are located in two distinct antigenic regions of the procolipase molecule. It appears then that the lipid binding domain of the pancreatic lipase protein cofactor comprises two regions. The first region corresponds to the amino terminal fragment of the protein. The second region is likely identical with the peptide segment at position 51-59 as previously hypothesized from NMR and spectrophotometric studies. Studies carried out on procolipase chemically modified at tyrosine residues provided evidence that epitopes for Mabs 49.20 and 75.8 are in or close to the region which contains tyrosines at positions 55 and 59, and that the two peptide regions essential for interfacial binding are spatially adjacent in the procolipase and the trypsin-treated form of the cofactor. General conclusions are in accordance with the location of antigenic regions of procolipase determined by predictive methods.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0006-3002
pubmed:author
pubmed:issnType
Print
pubmed:day
5
pubmed:volume
998
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
158-66
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:2477068-Animals, pubmed-meshheading:2477068-Antibodies, Monoclonal, pubmed-meshheading:2477068-Antibody Specificity, pubmed-meshheading:2477068-Binding, Competitive, pubmed-meshheading:2477068-Colipases, pubmed-meshheading:2477068-Enzyme Activation, pubmed-meshheading:2477068-Enzyme Precursors, pubmed-meshheading:2477068-Enzyme-Linked Immunosorbent Assay, pubmed-meshheading:2477068-Epitopes, pubmed-meshheading:2477068-Immunoglobulin Fab Fragments, pubmed-meshheading:2477068-Lipid Metabolism, pubmed-meshheading:2477068-Protein Conformation, pubmed-meshheading:2477068-Protein Precursors, pubmed-meshheading:2477068-Proteins, pubmed-meshheading:2477068-Species Specificity, pubmed-meshheading:2477068-Swine, pubmed-meshheading:2477068-Trypsin, pubmed-meshheading:2477068-Tyrosine
pubmed:year
1989
pubmed:articleTitle
Inhibitory properties and antigenic specificity of monoclonal antibodies to pancreatic colipase.
pubmed:affiliation
Laboratoire de Biochimie, Université de Provence, Marseille, France.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't