Structure of the pancreatic lipase–procolipase complex

Tilbeurgh, Herman van; Sarda, Louis; Verger, Robert; Cambillau, Christian

doi:10.1038/359159a0

Cited by 356 publications

(228 citation statements)

References 14 publications

Supporting

Mentioning

210

Contrasting

Unclassified

Order By: Relevance

“…Clustering of the hydrophobic residues (red) in the core of LCAT, due to the packing of the amphipathic helices against the hydrophobic residues of the P-sheet, and distribution of hydrophilic residues at the surface of the molecule, are illustrated on the right part of Figure 7. The residues of the catalytic triad are in the typical configuration of the alp hydrolase fold; the distance between the y 0 of SI81 and the N2 of H377 is 2.5 A, while that between 6 0 of D345 and the N1 of H377 is 2.9 A, comparable to the distances measured in crystallized lipases (Van Tilbeurgh et al, 1992. The backbone amides of the two oxyanion hole residues F103 and L182 involved in catalysis are located within 5 A of each other.…”

Section: Model Building and Biological Implicationssupporting

confidence: 59%

“…As this residue is located on the hydrophilic loop N-terminal of helix a4-5, the E149A mutation probably facilitates the spatial accommodation of a more bulky arachidonic acid molecule in this cavity. Fatty acid binding to cutinase (Longhi et al, 1996), to Rhizomucor miehei lipase (Vase1 et al, 1993;Norin et al, 1994), and to human pancreatic lipase (Van Tilbeurgh et al, 1992), involves similar interactions with a residue N-terminal of helix a4-5. The accommodation of the cholesterol nucleus in LCAT can be predicted from the structural and functional homology between LCAT and Candida cylindracea cholesterol esterase (Ghosh et al, 1995).…”

Section: Model Building and Biological Implicationsmentioning

confidence: 99%

See 1 more Smart Citation

A proposed architecture for lecithin cholesterol acyl transferase (LCAT): Identification of the catalytic triad and molecular modeling

et al. 1998

View full text Add to dashboard Cite

The enzyme cholesterol lecithin acyl transferase (LCAT) shares the Ser/Asp‐Glu/His triad with lipases, esterases and proteases, but the low level of sequence homology between LCAT and these enzymes did not allow for the LCAT fold to be identified yet. We, therefore, relied upon structural homology calculations using threading methods based on alignment of the sequence against a library of solved three‐dimensional protein structures, for prediction of the LCAT fold. We propose that LCAT, like lipases, belongs to the α/β hydrolase fold family, and that the central domain of LCAT consists of seven conserved parallel beta‐strands connected by four α‐helices and separated by loops. We used the conserved features of this protein fold for the prediction of functional domains in LCAT, and carried out site‐directed mutagenesis for the localization of the active site residues. The wild‐type enzyme and mutants were expressed in Cos‐1 cells. LCAT mass was measured by ELISA, and enzymatic activity was measured on recombinant HDL, on LDL and on a monomelic substrate. We identified D345 and H377 as the catalytic residues of LCAT, together with F103 and L182 as the oxyanion hole residues. In analogy with lipases, we further propose that a potential “lid” domain at residues 50‐74 of LCAT might be involved in the enzyme‐substrate interaction. Molecular modeling of human LCAT was carried out using human pancreatic and Candida antarctica lipases as templates. The three‐dimensional model proposed here is compatible with the position of natural mutants for either LCAT deficiency or Fish‐eye disease. It enables moreover prediction of the LCAT domains involved in the interaction with the phospholipid and cholesterol substrates.

show abstract

Section: Model Building and Biological Implicationssupporting

confidence: 59%

Section: Model Building and Biological Implicationsmentioning

confidence: 99%

A proposed architecture for lecithin cholesterol acyl transferase (LCAT): Identification of the catalytic triad and molecular modeling

et al. 1998

View full text Add to dashboard Cite

show abstract

“…The structures of colipase reported at 3.0 A resolution reveal no important differences with the present higher resolution model (van Tilbeurgh et al, 1992(van Tilbeurgh et al, , 1993a. Colipase is a flattened molecule of overall dimensions 25 x 30 X 35 A (Kinemage l).…”

Section: Secondary and Tertiary Structurecontrasting

confidence: 45%

“…The three regions consisting of the tips of the finger-like loops (residues 31-35, residues 51-55, and residues 71-79) have the highest B-factors and are the less well-defined areas within the molecule. Meanwhile, the definition of the electron density in these regions is much better than that of the 3.0-A-resolution structure of the closed pancreatic lipase-colipase complex (van Tilbeurgh et al, 1992) and it makes it possible for us to improve the quality of our model. Figure 3A and B show the 2F0 -F, residual electron density map for a well-defined (residue 43-48) and for a less well-defined region (residues 53-56).…”

Section: Quality Of the Structurementioning

confidence: 99%

“…The recently determined X-ray structures of pancreatic lipasecolipase complexes have shown that colipase binds exclusively to the noncatalytic C-terminal domain of pancreatic lipase in the absence of substrates or inhibitors (van Tilbeurgh et al, 1992(van Tilbeurgh et al, , 1993a. Upon binding of substrate, two surface loops of pancreatic lipase covering the active site undergo drastic conformational changes.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Crystallographic study of the structure of colipase and of the interaction with pancreatic lipase

Egloff

Sarda

Verger³

et al. 1995

Protein Science

Self Cite

View full text Add to dashboard Cite

Colipase ( M , 10 kDa) confers catalytic activity to pancreatic lipase under physiological conditions (high bile salt Colipase lacks well-defined secondary structure elements. This small protein seems to be stabilized mainly by an extended network of five disulfide bridges that runs throughout the flatly shaped molecule, reticulating its four finger-like loops. The colipase surface can be divided into a rather hydrophilic part, interacting with lipase, and a more hydrophobic part, formed by the tips of the fingers. The interaction between colipase and the C-terminal domain of lipase is stabilized by eight hydrogen bonds and about 80 van der Waals contacts. Upon opening of the lid, three more hydrogen bonds and about 28 van der Waals contacts are added, explaining the higher apparent affinity in the presence of a lipid/water interface. The tips of the fingers are very mobile and constitute the lipid interaction surface. Two detergent molecules that interact with colipase were observed in the crystal, covering part of the hydrophobic surface.

show abstract

Reactivation of the totally inactive pancreatic lipase RP1 by structure-predicted point mutations

Roussel

Bezzine

et al. 1998

Proteins

Self Cite

View full text Add to dashboard Cite

Both classical pancreatic lipase (DPL) and pancreatic lipase-related protein 1 (DPLRP1) have been found to be secreted by dog exocrine pancreas. These two proteins were purified to homogeneity from canine pancreatic juice and no significant catalytic activity was observed with dog PLRP1 on any of the substrates tested: di- and tri-glycerides, phospholipids, etc. DPLRP1 was crystallized and its structure solved by molecular replacement and refined at a resolution of 2.10 A. Its structure is similar to that of the classical PL structures in the absence of any inhibitors or micelles. The lid domain that controls the access to the active site was found to have a closed conformation. An amino-acid substitution (Ala 178 Val) in the DPLRP1 may result in a steric clash with one of the acyl chains observed in the structures of a C11 alkyl phosphonate inhibitor, a transition state analogue, bound to the classical PL. This substitution was suspected of being responsible for the absence of DPLRP1 activity. The presence of Val and Ala residues in positions 178 and 180, respectively, are characteristic of all the known PLRP1, whereas Ala and Pro residues are always present in the same positions in all the other members of the PL gene family. Introducing the double mutation Val 178 Ala and Ala 180 Pro into the human pancreatic RP1 (HPLRP1) gene yielded a well expressed and folded enzyme in insect cells. This enzyme is kinetically active on triglycerides. Our findings on DPLRP1 and HPLRP1 are therefore likely to apply to all the RP1 lipases.

show abstract

Structure of the pancreatic lipase–procolipase complex

Cited by 356 publications

References 14 publications

A proposed architecture for lecithin cholesterol acyl transferase (LCAT): Identification of the catalytic triad and molecular modeling

A proposed architecture for lecithin cholesterol acyl transferase (LCAT): Identification of the catalytic triad and molecular modeling

Crystallographic study of the structure of colipase and of the interaction with pancreatic lipase

Reactivation of the totally inactive pancreatic lipase RP1 by structure-predicted point mutations

Contact Info

Product

Resources

About