A small hydrophobic domain that localizes human erythrocyte acetylcholinesterase in liposomal membranes is cleaved by papain digestion

B. H. Kim, T. L. Rosenberry

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

A small hydrophobic domain in isolated human erythrocyte acetylcholinesterase is responsible for the interaction of this enzyme with detergent micelles and the aggregation of the enzyme on removal of detergent. Papain has been shown to cleave this hydrophobic domain and to generate a fully active hydrophilic enzyme that shows no tendency to interact with detergents or to aggregate [Dutta-Choudhury, T. A., & Rosenberry, T. L. (1984) J.Biol. Chem, 259, 5653-5660]. We report here that the intact enzyme could be reconstituted into phospholipid liposomes while the papain-disaggregated enzyme showed no capacity for reconstitution. More than 80% of the enzyme reconstituted into small liposomes could be released by papain digestion as the hydrophilic form. Papain was less effective in releasing the enzyme from large liposomes that were probably multilamellar. In a novel application of affinity chromatography on acridinium resin, enzyme reconstituted into small liposomes in the presence of excess phospholipid was purified to a level of 1 enzyme molecule per 4000 phospholipid molecules, a ratio expected if each enzyme molecule was associated with a small, unilamellar liposome. Subunits in the hydrophilic enzyme form released from reconstituted liposomes by papain digestion showed a mass decrease of about 2 kilodaltons relative to the intact subunits according to acrylamide gel electrophoresis in sodium dodecyl sulfate, a difference similar to that observed previously following papain digestion of the soluble enzyme aggregates. The data were consistent with the hypothesis that the same hydrophobic domain in the enzyme is responsible for the interaction of the enzyme with detergent micelles, the aggregation of the enzyme in the absence of detergent and the incorporation of the enzyme into reconstituted phospholipid membranes.

Original languageEnglish (US)
Pages (from-to)3586-3592
Number of pages7
JournalBiochemistry
Volume24
Issue number14
StatePublished - 1985
Externally publishedYes

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Papain
Acetylcholinesterase
Digestion
Erythrocytes
Membranes
Enzymes
Liposomes
Detergents
Phospholipids
Micelles
Molecules
Agglomeration
Affinity chromatography
Unilamellar Liposomes
Acrylamide
Electrophoresis
Affinity Chromatography
Sodium Dodecyl Sulfate

ASJC Scopus subject areas

  • Biochemistry

Cite this

A small hydrophobic domain that localizes human erythrocyte acetylcholinesterase in liposomal membranes is cleaved by papain digestion. / Kim, B. H.; Rosenberry, T. L.

In: Biochemistry, Vol. 24, No. 14, 1985, p. 3586-3592.

Research output: Contribution to journalArticle

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