The hydrophobic surface of colipase influences lipase activity at an oil-water interface.
The interaction of pancreatic triglyceride lipase and colipase at an oil-water interface is required for efficient digestion of dietary fats and provides a model system for the interaction of proteins at biological membranes. Colipase has two important surfaces, a hydrophilic surface that interacts with lipase and a hydrophobic surface that presumably interacts with substrate. To begin our investigations into the role of the hydrophobic surface in the function of colipase, we replaced three neighboring tyrosine residues at positions 55, 58, and 59 in the hydrophobic surface with aspartic acid. Two of the three residues, Tyr55 and Tyr59, influenced the activity of colipase. Introducing aspartic acid at either position decreased the activity with long-chain triglycerides, but not with a short-chain triglyceride. Decreased ability of the mutants to anchor lipase to long-chain triglycerides did not explain the altered activity of the mutants. A mutant containing aspartic acid at positions 55 and 59 had no activity with any substrate and did not anchor lipase to either short- or long-chain triglycerides. These results identify the two tyrosine residues that interact with substrate and suggest that the hydrophobicity of the surface containing these tyrosines influences colipase function and the substrate selectivity of pancreatic triglyceride lipase.
Cordle
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RA
Department of Pediatrics, Washington University School of Medicine, St. Louis, MO 63110, USA.
Lowe
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ME
D001224:Aspartic Acid; D002462:Cell Membrane / Q000201:enzymology; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003089:Colipases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D005217:Fat Emulsions, Intravenous / Q000378:metabolism; D006801:Humans; D008049:Lipase / Q000378:metabolism*; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D009821:Oils; D013329:Structure-Activity Relationship; D013499:Surface Properties; D014280:Triglycerides / Q000378:metabolism; D014304:Triolein / Q000378:metabolism; D014443:Tyrosine / Q000235:genetics; D014867:Water
D003089:Colipases; D005217:Fat Emulsions, Intravenous; D009821:Oils; D014280:Triglycerides; D014867:Water; D014304:Triolein; D001224:Aspartic Acid; D014443:Tyrosine; D008049:Lipase; C005830:tributyrin