Linked Life Data

8993339

Source:http://linkedlifedata.com/resource/pubmed/id/8993339

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
1997-2-5
pubmed:abstractText
We designed chimeric mutants by exchanging the lid domains of the classical human pancreatic lipase (HPL) and the guinea pig pancreatic lipase related protein 2 (GPLRP2). This latter enzyme possesses naturally a large deletion within the lid domain and is not activated by lipid/water interfaces. Furthermore, GPLRP2 exhibits phospholipase A1 and lipase activities in the same order of magnitude, whereas HPL has no significant phospholipase activity and displays a clear interfacial activation. An HPL mutant [HPL(-lid)] with GPLRP2 mini-lid domain does not display interfacial activation. Its specific activity toward triglycerides is, however, dramatically reduced. A GPLRP2 mutant [GPLRP2(+lid)] with HPL full-length lid domain is not interfacially activated, and its lid domain probably exists under a permanent open conformation. Therefore, the phenomenon of interfacial activation in HPL is not only due to the presence of a full-length lid domain but also to other structural elements which probably allow the existence of stabilized closed and open conformations of the lid. GPLRP2(+lid) phospholipase activity is significantly reduced as compared to GPLRP2, whereas its lipase activity remains at the same level. Therefore, the lid domain plays a major role in substrate selectivity and can be considered as part of the active site. However, the presence of a full-length lid domain is not sufficient to explain the absence of phospholipase activity in HPL since HPL(-lid) does not display any phospholipase activity. We also produced a chimeric GPLRP2 mutant in which the C-terminal domain was substituted by the HPL C-terminal domain. The colipase effects, i.e., anchoring and stabilization of the lipase at the interface, are clearly observed with the chimera, whereas GPLRP2 is insensitive to colipase. The kinetic characterization of this chimera reveals for the first time that the interfacial stability of pancreatic lipases depends on the structure of the C-terminal domain.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
7
pubmed:volume
36
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
239-48
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:8993339-Amino Acid Sequence, pubmed-meshheading:8993339-Animals, pubmed-meshheading:8993339-Base Sequence, pubmed-meshheading:8993339-Bile Acids and Salts, pubmed-meshheading:8993339-Colipases, pubmed-meshheading:8993339-DNA Primers, pubmed-meshheading:8993339-Enzyme Activation, pubmed-meshheading:8993339-Glycosylation, pubmed-meshheading:8993339-Guinea Pigs, pubmed-meshheading:8993339-Humans, pubmed-meshheading:8993339-Lipase, pubmed-meshheading:8993339-Molecular Sequence Data, pubmed-meshheading:8993339-Mutagenesis, Site-Directed, pubmed-meshheading:8993339-Mutation, pubmed-meshheading:8993339-Pancreas, pubmed-meshheading:8993339-Phospholipids, pubmed-meshheading:8993339-Recombinant Fusion Proteins, pubmed-meshheading:8993339-Sequence Analysis, pubmed-meshheading:8993339-Structure-Activity Relationship, pubmed-meshheading:8993339-Triglycerides
pubmed:year
1997
pubmed:articleTitle
Pancreatic lipase structure-function relationships by domain exchange.
pubmed:affiliation
Laboratoire de Lipolyse Enzymatique, UPR 9025, IFRI du CNRS, Marseille, France.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't