title: Identification of rat pancreatic secretory proteins after separation by high-performance liquid chromatography.
abstract: In the present study an improved method of reversed-phase high-performance liquid chromatography (HPLC) for separation of rat pancreatic juice proteins is introduced. Aliquots of pancreatic juice were saved from conscious rats during basal secretion. The secretory proteins were separated on a wide-pore silica column by use of a multistep acetonitrile/water gradient. Up to 14 individual peaks could be separated by one run. Molecular weight analysis by sodium dodecyl sulfate (SDS)-gels allowed identification of peaks representing amylase, lipase, procarboxypeptidases, proelastase, chymotrypsinogen, and trypsinogen. Injection of pure rat amylase increased one specific peak which was assumed to represent amylase in the juice profile. Small amounts of residual enzymatic activities were measured for amylase, trypsin, and chymotrypsin in material of certain peaks. Activities of lipase, ribonuclease, and carboxypeptidases were not found, which reflected degradation of these enzymes by the separation procedure. High activities of phospholipase A2 were detected in one specific, early-eluting peak. Reversed-phase HPLC offers precise, reproducible, and rapid separation of the major proteins of rat pancreatic juice.
authors: Department of Internal Medicine, Philipps University of Marburg, Federal Republic of Germany.
authors: B
authors: B
authors: Göke
authors: V
authors: V
authors: Keim
authors: T
authors: T
authors: Meyer
authors: R
authors: R
authors: Arnold
authors: G
authors: G
authors: Adler
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004792:Enzyme Precursors / Q000032:analysis*; D005260:Female; D006867:Hydrolases / Q000032:analysis*; D010189:Pancreatic Juice / Q000201:enzymology*; D010741:Phospholipases A / Q000032:analysis; D054467:Phospholipases A2; D051381:Rats; D011919:Rats, Inbred Strains; D014357:Trypsin / Q000032:analysis
chemical_list: D004792:Enzyme Precursors; D006867:Hydrolases; D010741:Phospholipases A; D054467:Phospholipases A2; D014357:Trypsin