Separation and analysis of pig pancreatic zymogen granules with free flow electrophoresis and lectins.
Purified pig pancreatic zymogen granules were subjected to free flow electrophoresis (FFE) in an acetate buffer system (acetic acid/NaOH, pH 5.5) to detect the presence or absence of more than one population or zymogen granules. Pig pancreatic zymogen granules were purified by differential and density gradient centrifugation and subjected to FFE. Fractions were analyzed for protein, alpha-amylase (EC 3.2.1.1) and 5'-nucleotidase (EC 3.1.3.5) as marker enzymes for zymogen granule content and membranes, respectively. Only one distinct peak, with coincident alpha-amylase and 5'-nucleotidase activity, and most protein was detected, which reflects the presence of a single population of intact zymogen granules. This was confirmed by electron microscopy. When the granules were incubated with different lectins before FFE, the one distinct peak representing intact zymogen granules was shifted towards the cathode in the case of concanavalin A (Con A) and Ricinus communis agglutinin 120 (RCA 120). No splitting of the peak occurred. Our results do not support the hypothesis of a coexistence of more than one distinct population of zymogen granules.
Spaans
M C
MC
Department of Internal Medicine, University Hospital, Zürich, Switzerland.
Tobler
M
M
Ammann
R W
RW
Freiburghaus
A U
AU
D000818:Animals; D001665:Binding Sites / Q000502:physiology; D003594:Cytoplasmic Granules / Q000378:metabolism*; D004586:Electrophoresis / Q000379:methods*; D004792:Enzyme Precursors / Q000302:isolation & purification*; D037102:Lectins / Q000378:metabolism*; D008854:Microscopy, Electron; D010179:Pancreas / Q000201:enzymology*; D013552:Swine
D004792:Enzyme Precursors; D037102:Lectins