Two-dimensional gel analysis of soluble proteins. Charaterization of guinea pig exocrine pancreatic proteins

J Biol Chem. 1975 Jul 25;250(14):5375-85.

Abstract

A two-dimensional gel technique using slab gel isoelectric focusing in the first dimension and sodium dodecyl sulfate gradient gel electrophoresis in the second dimension has been developed for the separation of soluble proteins larger than 10,000 daltons. The technique is sensitive to 0.6 mug of protein and recovery of radiolabeled proteins averages 90%. Analysis of secretory protein from the guinea pig exocrine pancreas shows the presence of 19 distinct high molecular weight proteins. Each of these proteins has been characterized by isoelectric point, molecular weight, and proportionate mass. Thirteen of the 19 proteins have been identified by actual or potential enzymatic activity,accounting for 96% of the protein mass resolved by the two-dimensional gels.

MeSH terms

  • Amino Acids
  • Amylases / metabolism
  • Animals
  • Autoradiography
  • Carbachol / pharmacology
  • Carbon Radioisotopes
  • Carboxypeptidases / metabolism
  • Chymotrypsinogen / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Precursors / metabolism
  • Female
  • Guinea Pigs
  • Isoelectric Focusing
  • Isotope Labeling
  • Lipase / metabolism
  • Molecular Weight
  • Pancreas / metabolism*
  • Pancreatic Elastase / metabolism
  • Proteins* / metabolism
  • Ribonucleases / metabolism
  • Sodium Dodecyl Sulfate
  • Trypsinogen / metabolism

Substances

  • Amino Acids
  • Carbon Radioisotopes
  • Enzyme Precursors
  • Proteins
  • Sodium Dodecyl Sulfate
  • Carbachol
  • Trypsinogen
  • Chymotrypsinogen
  • Ribonucleases
  • Lipase
  • Amylases
  • Carboxypeptidases
  • Pancreatic Elastase