Pyrularia Thionin Binding to and the Role of Tryptophan-8 in the Enhancement of Phosphatidylserine Domains in Erythrocyte Membranes

Fei Wang, Gary H. Naisbitt, Leo P. Vernon, Michael Glaser

Research output: Contribution to journalArticle

Abstract

Pyrularia thionin is a small, strongly basic peptide which interacts readily with cellular and synthetic membranes. With cells it induces hemolysis, depolarizes the cellular membrane with an accompanying influx of Ca2+, and activates an endogenous phospholipase A2. Evidence points toward a binding site involving phosphatidylserine (PS). This study shows that addition of the peptide to erythrocyte membranes as well as to vesicles formed from phospholipids isolated from erythrocyte membranes causes an enhancement of phospholipid domains which are made visible by the use of fluorescence digital imaging microscopy with fluorescent derivatives of PS (NBD-PS) and phosphatidylcholine (NBD-PC). Addition of thionin caused a large increase in NBD-PS domains, with an accompanying enrichment of NBD-PC in another separate domain. Double-labeling experiments performed with a Texas Red derivative of thionin show that the peptide binds to the domain enriched in NBD-PS. P thionin inactivated by modification of Trp-8 with N-bromosuccinimide lost the ability to enhance PS domains, although it bound to the membrane with the same affinity as native P thionin. This shows that binding to the membrane is not in itself sufficient to cause the NBD-PS and NBD-PC redistribution into domains.

Original languageEnglish (US)
Pages (from-to)12283-12289
Number of pages7
JournalBiochemistry
Volume32
Issue number46
DOIs
StatePublished - Jan 1 1993

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Pyrularia
Thionins
Phosphatidylserines
Erythrocyte Membrane
Tryptophan
Membranes
Peptides
Phospholipids
Bromosuccinimide
Derivatives
Phospholipases A2
Optical Imaging
Hemolysis
Phosphatidylcholines
Labeling
Microscopy
Microscopic examination
Fluorescence
Binding Sites

ASJC Scopus subject areas

  • Biochemistry

Cite this

Pyrularia Thionin Binding to and the Role of Tryptophan-8 in the Enhancement of Phosphatidylserine Domains in Erythrocyte Membranes. / Wang, Fei; Naisbitt, Gary H.; Vernon, Leo P.; Glaser, Michael.

In: Biochemistry, Vol. 32, No. 46, 01.01.1993, p. 12283-12289.

Research output: Contribution to journalArticle

Wang, Fei ; Naisbitt, Gary H. ; Vernon, Leo P. ; Glaser, Michael. / Pyrularia Thionin Binding to and the Role of Tryptophan-8 in the Enhancement of Phosphatidylserine Domains in Erythrocyte Membranes. In: Biochemistry. 1993 ; Vol. 32, No. 46. pp. 12283-12289.
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