Summary
Phospholipase A (PLA) is able to attack membrane phospholipids and thereby plays a putative role in the pathogenesis of pancreatic pseudocysts. We looked for PLA2-like activity in aspirates from human pancreatic pseudocysts. In material originating from one cyst which occurred shortly after an acute pancreatitis attack, hydrolyzing enzymatic activity measured by a sensitive bioassay system for PLA2 activity was found without prior trypsin activation (67×103 U/min/100 µl). A biochemical characterization of this hydrolyzing enzymatic activity was provided after resolution of the respective proteins contained in the cyst fluid by HPLC. High hydrolyzing activities were found in correspondence to one specific, early eluting peak. The purified enzyme had pH optima at 3.5 and 6. Addition of EDTA (5 mM) to the test system abolished the enzymatic activity which mirrored the requirement for calcium ions. The activity was optimal at calcium concentrations ranging from 1–2 mM. Higher calcium concentrations reduced the enzymatic activity. The enzyme showed high heat stability. SDS-gel analysis of the peak showed one single band with a molecular weight of about 20,000 Daltons. Our findings demonstrate the possibility of activated, PLA-like activity in human pancreatic pseudocyst fluid. We speculate that an inappropriate activation of this enzyme in peri- or intrapancreatic “fluid collections” could account for pseudocyst formation after an acute pancreatitis attack.
Similar content being viewed by others
References
Aho HJ, Nevalainen TJ, Lindenberg RLP, Aho HJ (1980) Experimental pancreatitis in the rat: the role of phospholipase A in sodium taurocholate-induced acute hemorrhagic pancreatitis. Scand J Gastroenterol 15:1027–1031
Bradley EL (1982) Pancreatic pseudocysts. In: Bradley EL (ed) Complications of pancreatitis: medical and surgical management. Saunders, Philadelphia
De Haas GH, Postema NH, Nieuwenhuizen W, Van Deneen LLM (1968) Purification and properties of an anionic zymogen of phospholipase A from porcine pancreas. Biochim Biophys Acta 159:118–129
De Haas GH, Slotboom AJ, Bonsen PPM, Van Deneen LLM (1970) Studies on phospholipase A and its zymogen from porcine pancreas: the complete amino acid sequence. Biochim Biophys Acta 221:31–53
Eskola JU, Nevalainen TJ, Aho HJ (1983) Purification and characterization of human pancreatic phospholipase A2. Clin Chem 29:1772–1776
Figarella C, Clemente F, Guy O (1971) A zymogen of phospholipase A in human pancreatic juice. Biochim Biophys Acta 227:213–217
Göke B, Keim V, Meyer T, Arnold R, Adler G (1988) Identification of rat pancreatic secretory proteins after separation by high performance liquid chromatography. Pancreas 3:199–206
Grataroli R, Dijkman R, Dutilh CE, Van der Ondera F, De Haas GH, Figarella C (1982) Studies on phospholipase A2 and its enzyme from human pancreatic juice. Eur J Biochem 122:111–117
Laemmli U, Favre M (1973) Maturation of head bacteriophage. J Mol Biol 80:525–599
Lasson A, Ohlsson K (1987) Pancreatic pseudocysts: a biochemical evaluation of proteases and protease inhibitors in plasma. Scand J Gastroenterol 22:355–361
Maule WF, Reber HA (1986) Diagnosis and management of pancreatic pseudocysts, pancreatic ascites, and pancreatic fistulas. In: Go VWL, Gardner JD, Brooks FP, Lebenthal E, DiMagno EP, Scheele GA (eds) The exocrine pancreas: biology, pathobiology, and diseases. Raven Press, New York, pp 601–610
Nevalainen TJ (1980) The role of phospholipase A in acute pancreatitis. Scand J Gastroenterol 15:641–650
Nevalainen TJ, Aho HJ, Eskola JU, Suonpaa AK (1983) Immunohistochemical localization of phospholipase A in human pancreas in acute and chronic pancreatitis. Acta Pathol Microbiol Scand (A) 91:97–102
Patriarca P, Beckerdite S, Elsbach P (1972) Phospholipases and physiological turnover inEschericia coli spheroblasts. Biochim Biophys Acta 260:593–600
Rinderknecht H (1986) Pancreatic secretory enzymes. In: Go VWL, Gardner JD, Brooks FP, Lebenthal E, DiMagno EP, Scheele GA (eds) The exocrine pancreas: biology, pathobiology, and diseases. Raven Press, New York, pp 163–183
Schröder T, Somerharjn P, Lempinen M (1979) Phospholipase A in serum and ascitic exudate in experimental acute hemorrhagic pancreatitis in pigs. Eur Surg Res 11:172–178
Tagesson C, Sjödahl R (1985) Studies of the phospholipase A2 activity of rat ileal mucosa. Scand J Gastroenterol 20:25–30
Vadas P (1982) The efficiancy of anti-inflammatory agents with respect to extracellular phospholipase A2 activity. Life Sci 30:155–162R
Vadas P, Hay JB (1980) The release of phospholipase A2 from aggregated platelets and stimulated macrophages of sheep. Life Sci 26:1721–1729
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Göke, B., Meyer, T., Loth, H. et al. Characterization of phospholipase A2 activity in aspirates of human pancreatic pseudocysts after isolation by reversed-phase high performance liquid chromatography. Klin Wochenschr 67, 131–135 (1989). https://doi.org/10.1007/BF01711338
Issue Date:
DOI: https://doi.org/10.1007/BF01711338