High density sickle cell erythrocyte core membrane skeletons demonstrate slow temperature dependent dissociation

Archil Shartava, Paul Miranda, Kenya N. Williams, Arvind Shah, Carlos A. Monteiro, Steven Goodman

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

We have previously demonstrated that slow dissociation of HDSS membrane skeletons in high ionic strength Triton X-100 buffer was related to a posttranslational modification in β-actin, in which a disulfide bridge was formed between cysteine 284 and cysteine 373 [Shartava et al: J Cell Biol 128:805, 1995]. These previous dissociation assays were limited to two homozygous (SS) sickle cell patients and a single temperature (37°C). In the current work, we have expanded the SS subjects to 9 and have carried out dissociation assays at 0, 24, 30, 34, and 37°C. At 0°C there was limited dissociation of spectrin and actin from normal (AA), low density sickle cell (LDSS), and high density sickle cell (HDSS) core skeleton up to 24 hr. The first order rate constants for dissociation of spectrin, at 0°C, was 0.030-0.035 x 10-4 sec-1 for AA, LDSS, and HDSS core skeletons. However at 24, 30, 34, and 37°C the rate of dissociation of spectrin from HDSS core skeletons was significantly slower than the rate of dissociation from AA core skeletons. Having determined the first order rate constants for spectrin dissociation at these specified temperatures, we then asked whether dithiothreitol (DTT) would hasten the dissociation of core skeletons. The presence of DTT caused the rate of dissociation of the HDSS membrane skeleton to become statistically indistinguishable from the rate of dissociation of AA membrane skeletons. This is consistent with the suggestion that reversible thiol oxidation is responsible for the slow dissociation of the HDSS membrane skeleton.

Original languageEnglish (US)
Pages (from-to)214-219
Number of pages6
JournalAmerican Journal of Hematology
Volume51
Issue number3
DOIs
StatePublished - Mar 1 1996
Externally publishedYes

Fingerprint

Erythrocyte Membrane
Skeleton
Cell Count
Temperature
Spectrin
Dithiothreitol
Cell Membrane
Cysteine
Actins
Octoxynol
Post Translational Protein Processing
Sulfhydryl Compounds
Disulfides
Osmolar Concentration
Buffers
Membranes

All Science Journal Classification (ASJC) codes

  • Hematology

Cite this

High density sickle cell erythrocyte core membrane skeletons demonstrate slow temperature dependent dissociation. / Shartava, Archil; Miranda, Paul; Williams, Kenya N.; Shah, Arvind; Monteiro, Carlos A.; Goodman, Steven.

In: American Journal of Hematology, Vol. 51, No. 3, 01.03.1996, p. 214-219.

Research output: Contribution to journalArticle

Shartava, Archil ; Miranda, Paul ; Williams, Kenya N. ; Shah, Arvind ; Monteiro, Carlos A. ; Goodman, Steven. / High density sickle cell erythrocyte core membrane skeletons demonstrate slow temperature dependent dissociation. In: American Journal of Hematology. 1996 ; Vol. 51, No. 3. pp. 214-219.
@article{bfab676748784f2aabf264462507cdfe,
title = "High density sickle cell erythrocyte core membrane skeletons demonstrate slow temperature dependent dissociation",
abstract = "We have previously demonstrated that slow dissociation of HDSS membrane skeletons in high ionic strength Triton X-100 buffer was related to a posttranslational modification in β-actin, in which a disulfide bridge was formed between cysteine 284 and cysteine 373 [Shartava et al: J Cell Biol 128:805, 1995]. These previous dissociation assays were limited to two homozygous (SS) sickle cell patients and a single temperature (37°C). In the current work, we have expanded the SS subjects to 9 and have carried out dissociation assays at 0, 24, 30, 34, and 37°C. At 0°C there was limited dissociation of spectrin and actin from normal (AA), low density sickle cell (LDSS), and high density sickle cell (HDSS) core skeleton up to 24 hr. The first order rate constants for dissociation of spectrin, at 0°C, was 0.030-0.035 x 10-4 sec-1 for AA, LDSS, and HDSS core skeletons. However at 24, 30, 34, and 37°C the rate of dissociation of spectrin from HDSS core skeletons was significantly slower than the rate of dissociation from AA core skeletons. Having determined the first order rate constants for spectrin dissociation at these specified temperatures, we then asked whether dithiothreitol (DTT) would hasten the dissociation of core skeletons. The presence of DTT caused the rate of dissociation of the HDSS membrane skeleton to become statistically indistinguishable from the rate of dissociation of AA membrane skeletons. This is consistent with the suggestion that reversible thiol oxidation is responsible for the slow dissociation of the HDSS membrane skeleton.",
author = "Archil Shartava and Paul Miranda and Williams, {Kenya N.} and Arvind Shah and Monteiro, {Carlos A.} and Steven Goodman",
year = "1996",
month = "3",
day = "1",
doi = "10.1002/(SICI)1096-8652(199603)51:3<214::AID-AJH6>3.0.CO;2-#",
language = "English (US)",
volume = "51",
pages = "214--219",
journal = "American Journal of Hematology",
issn = "0361-8609",
publisher = "Wiley-Liss Inc.",
number = "3",

}

TY - JOUR

T1 - High density sickle cell erythrocyte core membrane skeletons demonstrate slow temperature dependent dissociation

AU - Shartava, Archil

AU - Miranda, Paul

AU - Williams, Kenya N.

AU - Shah, Arvind

AU - Monteiro, Carlos A.

AU - Goodman, Steven

PY - 1996/3/1

Y1 - 1996/3/1

N2 - We have previously demonstrated that slow dissociation of HDSS membrane skeletons in high ionic strength Triton X-100 buffer was related to a posttranslational modification in β-actin, in which a disulfide bridge was formed between cysteine 284 and cysteine 373 [Shartava et al: J Cell Biol 128:805, 1995]. These previous dissociation assays were limited to two homozygous (SS) sickle cell patients and a single temperature (37°C). In the current work, we have expanded the SS subjects to 9 and have carried out dissociation assays at 0, 24, 30, 34, and 37°C. At 0°C there was limited dissociation of spectrin and actin from normal (AA), low density sickle cell (LDSS), and high density sickle cell (HDSS) core skeleton up to 24 hr. The first order rate constants for dissociation of spectrin, at 0°C, was 0.030-0.035 x 10-4 sec-1 for AA, LDSS, and HDSS core skeletons. However at 24, 30, 34, and 37°C the rate of dissociation of spectrin from HDSS core skeletons was significantly slower than the rate of dissociation from AA core skeletons. Having determined the first order rate constants for spectrin dissociation at these specified temperatures, we then asked whether dithiothreitol (DTT) would hasten the dissociation of core skeletons. The presence of DTT caused the rate of dissociation of the HDSS membrane skeleton to become statistically indistinguishable from the rate of dissociation of AA membrane skeletons. This is consistent with the suggestion that reversible thiol oxidation is responsible for the slow dissociation of the HDSS membrane skeleton.

AB - We have previously demonstrated that slow dissociation of HDSS membrane skeletons in high ionic strength Triton X-100 buffer was related to a posttranslational modification in β-actin, in which a disulfide bridge was formed between cysteine 284 and cysteine 373 [Shartava et al: J Cell Biol 128:805, 1995]. These previous dissociation assays were limited to two homozygous (SS) sickle cell patients and a single temperature (37°C). In the current work, we have expanded the SS subjects to 9 and have carried out dissociation assays at 0, 24, 30, 34, and 37°C. At 0°C there was limited dissociation of spectrin and actin from normal (AA), low density sickle cell (LDSS), and high density sickle cell (HDSS) core skeleton up to 24 hr. The first order rate constants for dissociation of spectrin, at 0°C, was 0.030-0.035 x 10-4 sec-1 for AA, LDSS, and HDSS core skeletons. However at 24, 30, 34, and 37°C the rate of dissociation of spectrin from HDSS core skeletons was significantly slower than the rate of dissociation from AA core skeletons. Having determined the first order rate constants for spectrin dissociation at these specified temperatures, we then asked whether dithiothreitol (DTT) would hasten the dissociation of core skeletons. The presence of DTT caused the rate of dissociation of the HDSS membrane skeleton to become statistically indistinguishable from the rate of dissociation of AA membrane skeletons. This is consistent with the suggestion that reversible thiol oxidation is responsible for the slow dissociation of the HDSS membrane skeleton.

UR - http://www.scopus.com/inward/record.url?scp=0029883813&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029883813&partnerID=8YFLogxK

U2 - 10.1002/(SICI)1096-8652(199603)51:3<214::AID-AJH6>3.0.CO;2-#

DO - 10.1002/(SICI)1096-8652(199603)51:3<214::AID-AJH6>3.0.CO;2-#

M3 - Article

VL - 51

SP - 214

EP - 219

JO - American Journal of Hematology

JF - American Journal of Hematology

SN - 0361-8609

IS - 3

ER -