In vitro lipolysis by human pancreatic lipase is specifically abolished by its inactive forms

Miled, Nabil; Berti-Dupuis, Liliane; Rivière, Mireille; Carrière, Frédéric; Verger, Robert

doi:10.1016/s1570-9639(02)00537-x

Cited by 5 publications

(4 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…3A, lane 5), show that the lipase-colipase interactions are strengthened in the case of the inhibited (open) HPL. This conclusion is in good agreement with our previous studies on the inhibition of HPL by its covalently inactivated form via a process of colipase sequestration [51].…”

Section: Discussionsupporting

confidence: 93%

Discrimination between closed and open forms of lipases using electrophoretic techniques

Miled¹,

Rivière²,

Cavalier³

et al. 2005

Analytical Biochemistry

View full text Add to dashboard Cite

Section: Discussionsupporting

confidence: 93%

Discrimination between closed and open forms of lipases using electrophoretic techniques

Miled¹,

Rivière²,

Cavalier³

et al. 2005

Analytical Biochemistry

View full text Add to dashboard Cite

“…The inhibitory effect was also observed on hydrolysis by PLRP2, only when colipase‐sensitive NP‐PLRP2 form was used. PLRP1 was reported to bind colipase in vitro 42, 43. We conclude that the effect of PLRP1 on milk fat globule hydrolysis by PL or NP‐PLRP2 is due to a competition for colipase binding.…”

Section: Discussionmentioning

confidence: 75%

Individual and combined action of pancreatic lipase and pancreatic lipase‐related proteins 1 and 2 on native versus homogenized milk fat globules

Berton

Sebban‐Kreuzer

Rouvellac

et al. 2009

Molecular Nutrition Food Res

View full text Add to dashboard Cite

Pancreatic lipase (PL) and pancreatic lipase-related proteins 1 and 2 (PLRP1 and PLRP2) display different functional properties, despite close structures. The aim of the study was to compare the kinetic properties of recombinant human PLRP1, PLRP2, and PL on a physiological substrate: the milk fat under native and homogenized structures. No lipolytic activity is measured for PLRP1. PLRP2 hydrolyses milk fat with a lower catalytic efficiency than that of PL. PLRP2 activity, higher on homogenized than on native milk fat, is differently influenced by fatty acids (FA) and colipase depending on a proteolytic cleavage in the lid domain. FA enhance the activity on both milks. A colipase positive effect on the non-proteolyzed PLRP2 is observed on homogenized milk and with FA on native milk fat. Bile salts are necessary. An original observation is a synergic effect between PL and PLRP2 on the two milks. An inhibitory effect of PLRP1 on PL activity is also demonstrated. The combined action of pancreatic lipases on milk fat digestion proposes PLRPs as modulators of PL. Our study supports the hypothesis of a major role of PLRP2 in fat digestion in newborns and brings new insights to understand the physiological role of PLRPs.

show abstract

“…Indeed, the [N2Cc]/[lipase] molar ratio needed to obtain 50% inhibition is the same as that found with E600‐hoPL competitor or with other inactive forms of PL used as competitors by Miled et al . [41]. This result indicates that N2Cc binds to colipase as well as hoPL.…”

Section: Discussionmentioning

confidence: 90%

Role of the structural domains in the functional properties of pancreatic lipase‐related protein 2

Berton¹,

Sebban‐Kreuzer²,

Crenon³

2007

The FEBS Journal

View full text Add to dashboard Cite

In 1992, Giller et al. isolated mRNA coding for two novel human pancreatic lipase-related proteins (PLRPs) showing a high level of identity with the human classic pancreatic lipase [1]. On the basis of amino acid sequence comparisons, Giller et al. proposed the classification of pancreatic lipases in three subgroups: classic pancreatic lipase (PL), PLRP1 and PLRP2. Numerous PLRP sequences have been identified in several species by isolating mRNA [2][3][4][5][6][7][8][9][10][11]. Furthermore, by using classic protein purification procedures, the presence of PLRP1 and ⁄ or PLRP2 has been demonstrated in the pancreas or in the pancreatic juice from different species and also in other secretions [8,[11][12][13][14][15].PLRP and PL differ in enzymatic properties such as substrate specificity, sensitivity to inhibition by bile salts and colipase dependence [16]. Pancreatic lipases are highly active and selective for triglyceride substrates. Under physiological conditions, the PL activity is dependent on the presence of colipase, which able to overcome the inhibitory effect of bile salts [17,18]. Despite extensive studies on a large variety of substrates, only very low lipolytic activity against triglycerides has been reported with PLRP1 [1,4,14,15] Although structurally similar, classic pancreatic lipase (PL) and pancreatic lipase-related protein (PLRP)2, expressed in the pancreas of several species, differ in substrate specificity, sensitivity to bile salts and colipase dependence. In order to investigate the role of the two domains of PLRP2 in the function of the protein, two chimeric proteins were designed by swapping the N and C structural domains between the horse PL (Nc and Cc domains) and the horse PLRP2 (N2 and C2 domains). NcC2 and N2Cc proteins were expressed in insect cells, purified by one-step chromatography, and characterized. NcC2 displays the same specific activity as PL, whereas N2Cc has the same as that PLRP2. In contrast to N2Cc, NcC2 is highly sensitive to interfacial denaturation. The lipolytic activity of both chimeric proteins is inhibited by bile salts and is not restored by colipase. Only N2Cc is found to be a strong inhibitor of PL activity, due to competition for colipase binding. Active site-directed inhibition experiments demonstrate that activation of N2Cc occurs in the presence of bile salt and does not require colipase, as does PLRP2. The inability of PLRP2 to form a high-affinity complex with colipase is only due to the C-terminal domain. Indeed, the N-terminal domain can interact with the colipase. PLRP2 properties such as substrate selectivity, specific activity, bile salt-dependent activation and interfacial stability depend on the nature of the N-terminal domain.

show abstract

In vitro lipolysis by human pancreatic lipase is specifically abolished by its inactive forms

Cited by 5 publications

References 33 publications

Discrimination between closed and open forms of lipases using electrophoretic techniques

Discrimination between closed and open forms of lipases using electrophoretic techniques

Individual and combined action of pancreatic lipase and pancreatic lipase‐related proteins 1 and 2 on native versus homogenized milk fat globules

Role of the structural domains in the functional properties of pancreatic lipase‐related protein 2

Contact Info

Product

Resources

About