Hemolytic properties under hydrostatic pressure of neuraminidase or protease-treated human erythrocytes

Takeo Yamaguchi, Masaki Matsumoto, Eiji Kimoto

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

We investigated the hemolytic properties under high pressure (200 MPa) of human erythrocytes in which sialic acids and glycopeptides had been removed from membrane surface by using neuraminidase and proteolytic enzymes such as trypsin and chymotrypsin, respectively. The degree of hemolysis increased in proportion to the amounts of sialic acids or glycopeptides released from intact erythrocytes. Studies of the time course of hemolysis showed that upon enzymatic digestion erythrocyte membranes became more fragile against high pressure. Such fragility decreased in the presence of chlorpromazine and trifluo-perazine but was unaffected by chlorpromazine methiodide or indomethacin. Furthermore, the effect of cross-linking of membrane proteins by diamide on the fragility was examined. The degree of hemolysis at 200 MPa increased upon removal of sialic acids from red cells in which spectrin is mainly cross-linked, but did not upon enzymatic digestion of red cells in which glycophorins, in addition to cross-linking of themselves, are included in the large-molecular-weight aggregates formed by cross-linking of the membrane skeleton with transmembrane proteins. In the latter case, however, upon reduction of the cross-linking by dithiothreitol the effect of enzymatic digestion appeared again. On the other hand, such an enzymatic digestion effect on osmotic hemolysis was not observed either in intact erythrocytes or in diamide-treated red cells. These results suggest that the interaction of the cytoplasmic domains of glycophorins with cytoskeletal proteins may be weakened by enzymatic digestion of the exofacial domains of glycophorins.

Original languageEnglish
Pages (from-to)576-581
Number of pages6
JournalJournal of biochemistry
Volume114
Issue number4
DOIs
Publication statusPublished - Jan 1 1993
Externally publishedYes

Fingerprint

Sialic Acids
Glycophorin
Hydrostatic Pressure
Neuraminidase
Hydrostatic pressure
Diamide
Digestion
Hemolysis
Peptide Hydrolases
Glycopeptides
Erythrocytes
Cells
Membranes
Perazine
Spectrin
Cytoskeletal Proteins
Dithiothreitol
Chlorpromazine
Indomethacin
Membrane Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Hemolytic properties under hydrostatic pressure of neuraminidase or protease-treated human erythrocytes. / Yamaguchi, Takeo; Matsumoto, Masaki; Kimoto, Eiji.

In: Journal of biochemistry, Vol. 114, No. 4, 01.01.1993, p. 576-581.

Research output: Contribution to journalArticle

@article{f7dc7f26d64d489b8bdc73a2484c750e,
title = "Hemolytic properties under hydrostatic pressure of neuraminidase or protease-treated human erythrocytes",
abstract = "We investigated the hemolytic properties under high pressure (200 MPa) of human erythrocytes in which sialic acids and glycopeptides had been removed from membrane surface by using neuraminidase and proteolytic enzymes such as trypsin and chymotrypsin, respectively. The degree of hemolysis increased in proportion to the amounts of sialic acids or glycopeptides released from intact erythrocytes. Studies of the time course of hemolysis showed that upon enzymatic digestion erythrocyte membranes became more fragile against high pressure. Such fragility decreased in the presence of chlorpromazine and trifluo-perazine but was unaffected by chlorpromazine methiodide or indomethacin. Furthermore, the effect of cross-linking of membrane proteins by diamide on the fragility was examined. The degree of hemolysis at 200 MPa increased upon removal of sialic acids from red cells in which spectrin is mainly cross-linked, but did not upon enzymatic digestion of red cells in which glycophorins, in addition to cross-linking of themselves, are included in the large-molecular-weight aggregates formed by cross-linking of the membrane skeleton with transmembrane proteins. In the latter case, however, upon reduction of the cross-linking by dithiothreitol the effect of enzymatic digestion appeared again. On the other hand, such an enzymatic digestion effect on osmotic hemolysis was not observed either in intact erythrocytes or in diamide-treated red cells. These results suggest that the interaction of the cytoplasmic domains of glycophorins with cytoskeletal proteins may be weakened by enzymatic digestion of the exofacial domains of glycophorins.",
author = "Takeo Yamaguchi and Masaki Matsumoto and Eiji Kimoto",
year = "1993",
month = "1",
day = "1",
doi = "10.1093/oxfordjournals.jbchem.a124219",
language = "English",
volume = "114",
pages = "576--581",
journal = "Journal of Biochemistry",
issn = "0021-924X",
publisher = "Oxford University Press",
number = "4",

}

TY - JOUR

T1 - Hemolytic properties under hydrostatic pressure of neuraminidase or protease-treated human erythrocytes

AU - Yamaguchi, Takeo

AU - Matsumoto, Masaki

AU - Kimoto, Eiji

PY - 1993/1/1

Y1 - 1993/1/1

N2 - We investigated the hemolytic properties under high pressure (200 MPa) of human erythrocytes in which sialic acids and glycopeptides had been removed from membrane surface by using neuraminidase and proteolytic enzymes such as trypsin and chymotrypsin, respectively. The degree of hemolysis increased in proportion to the amounts of sialic acids or glycopeptides released from intact erythrocytes. Studies of the time course of hemolysis showed that upon enzymatic digestion erythrocyte membranes became more fragile against high pressure. Such fragility decreased in the presence of chlorpromazine and trifluo-perazine but was unaffected by chlorpromazine methiodide or indomethacin. Furthermore, the effect of cross-linking of membrane proteins by diamide on the fragility was examined. The degree of hemolysis at 200 MPa increased upon removal of sialic acids from red cells in which spectrin is mainly cross-linked, but did not upon enzymatic digestion of red cells in which glycophorins, in addition to cross-linking of themselves, are included in the large-molecular-weight aggregates formed by cross-linking of the membrane skeleton with transmembrane proteins. In the latter case, however, upon reduction of the cross-linking by dithiothreitol the effect of enzymatic digestion appeared again. On the other hand, such an enzymatic digestion effect on osmotic hemolysis was not observed either in intact erythrocytes or in diamide-treated red cells. These results suggest that the interaction of the cytoplasmic domains of glycophorins with cytoskeletal proteins may be weakened by enzymatic digestion of the exofacial domains of glycophorins.

AB - We investigated the hemolytic properties under high pressure (200 MPa) of human erythrocytes in which sialic acids and glycopeptides had been removed from membrane surface by using neuraminidase and proteolytic enzymes such as trypsin and chymotrypsin, respectively. The degree of hemolysis increased in proportion to the amounts of sialic acids or glycopeptides released from intact erythrocytes. Studies of the time course of hemolysis showed that upon enzymatic digestion erythrocyte membranes became more fragile against high pressure. Such fragility decreased in the presence of chlorpromazine and trifluo-perazine but was unaffected by chlorpromazine methiodide or indomethacin. Furthermore, the effect of cross-linking of membrane proteins by diamide on the fragility was examined. The degree of hemolysis at 200 MPa increased upon removal of sialic acids from red cells in which spectrin is mainly cross-linked, but did not upon enzymatic digestion of red cells in which glycophorins, in addition to cross-linking of themselves, are included in the large-molecular-weight aggregates formed by cross-linking of the membrane skeleton with transmembrane proteins. In the latter case, however, upon reduction of the cross-linking by dithiothreitol the effect of enzymatic digestion appeared again. On the other hand, such an enzymatic digestion effect on osmotic hemolysis was not observed either in intact erythrocytes or in diamide-treated red cells. These results suggest that the interaction of the cytoplasmic domains of glycophorins with cytoskeletal proteins may be weakened by enzymatic digestion of the exofacial domains of glycophorins.

UR - http://www.scopus.com/inward/record.url?scp=0027439105&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027439105&partnerID=8YFLogxK

U2 - 10.1093/oxfordjournals.jbchem.a124219

DO - 10.1093/oxfordjournals.jbchem.a124219

M3 - Article

C2 - 8276771

AN - SCOPUS:0027439105

VL - 114

SP - 576

EP - 581

JO - Journal of Biochemistry

JF - Journal of Biochemistry

SN - 0021-924X

IS - 4

ER -