Lamellar body-enriched fractions from neonatal mice: Preparative techniques and partial characterization

S. Grayson, A. G. Johnson-Winegar, B. U. Wintroub, R. R. Isseroff, E. H. Epstein, P. M. Elias

Research output: Contribution to journalArticle

132 Citations (Scopus)

Abstract

Several problems have frustrated the isolation of lamellar bodies (LB) from mammalian epidermis. We obtained pellets enriched in intact LB by utilizing the staphylococcal epidermolytic toxin to provide intact, outer epidermal sheets, by controlled homogenization in a cell disrupter, and by passage of homogenates through a graded series of nuclepore filters (Science 221:962, 1983). Such preparations contained more intact LB than did fractions prepared by a variety of differential or sucrose/metrizamide discontinuous centrifugation methods. Initial characterization of the enzymatic content of this fraction revealed it to be enriched in certain hydrolytic enzymes (acid phosphatase, carboxypeptidase, cathepsin B, acid lipase, sphingomyelinase, and phospholipase A), but strikingly depleted in all sulfatases, β-glucuronidase, and the non-lysosomal protease, plasminogen activator. Thus, LB show some properties of lysosomes, although certain characteristic lysosomal enzymes are strikingly absent. Lamellar body fractions contained 2-3 times more lipid per unit weight than did homogenates, and were enriched in phospholipids, free sterols, and glycosphingolipids, but not in other neutral lipids or ceramides. In summary, whereas some of the enzymes in LB could participate in the metabolism of LB lipid precursors to hydrophobic barrier constituents, other may attack intercellular constituents, ultimately resulting in desquamation. The lipid profile of these organelles suggests that they deliver precursors of permeability barrier lipids to intercellular domains.

Original languageEnglish (US)
Pages (from-to)289-294
Number of pages6
JournalJournal of Investigative Dermatology
Volume85
Issue number4
StatePublished - 1985

Fingerprint

Lipids
Enzymes
Exfoliatins
Carboxypeptidase B
Cathepsin A
Metrizamide
Sulfatases
Sphingomyelin Phosphodiesterase
Carboxypeptidases
Glycosphingolipids
Cathepsin B
Centrifugation
Phospholipases A
Plasminogen Activators
Ceramides
Glucuronidase
Sterols
Acid Phosphatase
Lysosomes
Lipase

ASJC Scopus subject areas

  • Dermatology

Cite this

Grayson, S., Johnson-Winegar, A. G., Wintroub, B. U., Isseroff, R. R., Epstein, E. H., & Elias, P. M. (1985). Lamellar body-enriched fractions from neonatal mice: Preparative techniques and partial characterization. Journal of Investigative Dermatology, 85(4), 289-294.

Lamellar body-enriched fractions from neonatal mice : Preparative techniques and partial characterization. / Grayson, S.; Johnson-Winegar, A. G.; Wintroub, B. U.; Isseroff, R. R.; Epstein, E. H.; Elias, P. M.

In: Journal of Investigative Dermatology, Vol. 85, No. 4, 1985, p. 289-294.

Research output: Contribution to journalArticle

Grayson, S, Johnson-Winegar, AG, Wintroub, BU, Isseroff, RR, Epstein, EH & Elias, PM 1985, 'Lamellar body-enriched fractions from neonatal mice: Preparative techniques and partial characterization', Journal of Investigative Dermatology, vol. 85, no. 4, pp. 289-294.
Grayson, S. ; Johnson-Winegar, A. G. ; Wintroub, B. U. ; Isseroff, R. R. ; Epstein, E. H. ; Elias, P. M. / Lamellar body-enriched fractions from neonatal mice : Preparative techniques and partial characterization. In: Journal of Investigative Dermatology. 1985 ; Vol. 85, No. 4. pp. 289-294.
@article{8264403fe23c4f39857046220359583c,
title = "Lamellar body-enriched fractions from neonatal mice: Preparative techniques and partial characterization",
abstract = "Several problems have frustrated the isolation of lamellar bodies (LB) from mammalian epidermis. We obtained pellets enriched in intact LB by utilizing the staphylococcal epidermolytic toxin to provide intact, outer epidermal sheets, by controlled homogenization in a cell disrupter, and by passage of homogenates through a graded series of nuclepore filters (Science 221:962, 1983). Such preparations contained more intact LB than did fractions prepared by a variety of differential or sucrose/metrizamide discontinuous centrifugation methods. Initial characterization of the enzymatic content of this fraction revealed it to be enriched in certain hydrolytic enzymes (acid phosphatase, carboxypeptidase, cathepsin B, acid lipase, sphingomyelinase, and phospholipase A), but strikingly depleted in all sulfatases, β-glucuronidase, and the non-lysosomal protease, plasminogen activator. Thus, LB show some properties of lysosomes, although certain characteristic lysosomal enzymes are strikingly absent. Lamellar body fractions contained 2-3 times more lipid per unit weight than did homogenates, and were enriched in phospholipids, free sterols, and glycosphingolipids, but not in other neutral lipids or ceramides. In summary, whereas some of the enzymes in LB could participate in the metabolism of LB lipid precursors to hydrophobic barrier constituents, other may attack intercellular constituents, ultimately resulting in desquamation. The lipid profile of these organelles suggests that they deliver precursors of permeability barrier lipids to intercellular domains.",
author = "S. Grayson and Johnson-Winegar, {A. G.} and Wintroub, {B. U.} and Isseroff, {R. R.} and Epstein, {E. H.} and Elias, {P. M.}",
year = "1985",
language = "English (US)",
volume = "85",
pages = "289--294",
journal = "Journal of Investigative Dermatology",
issn = "0022-202X",
publisher = "Nature Publishing Group",
number = "4",

}

TY - JOUR

T1 - Lamellar body-enriched fractions from neonatal mice

T2 - Preparative techniques and partial characterization

AU - Grayson, S.

AU - Johnson-Winegar, A. G.

AU - Wintroub, B. U.

AU - Isseroff, R. R.

AU - Epstein, E. H.

AU - Elias, P. M.

PY - 1985

Y1 - 1985

N2 - Several problems have frustrated the isolation of lamellar bodies (LB) from mammalian epidermis. We obtained pellets enriched in intact LB by utilizing the staphylococcal epidermolytic toxin to provide intact, outer epidermal sheets, by controlled homogenization in a cell disrupter, and by passage of homogenates through a graded series of nuclepore filters (Science 221:962, 1983). Such preparations contained more intact LB than did fractions prepared by a variety of differential or sucrose/metrizamide discontinuous centrifugation methods. Initial characterization of the enzymatic content of this fraction revealed it to be enriched in certain hydrolytic enzymes (acid phosphatase, carboxypeptidase, cathepsin B, acid lipase, sphingomyelinase, and phospholipase A), but strikingly depleted in all sulfatases, β-glucuronidase, and the non-lysosomal protease, plasminogen activator. Thus, LB show some properties of lysosomes, although certain characteristic lysosomal enzymes are strikingly absent. Lamellar body fractions contained 2-3 times more lipid per unit weight than did homogenates, and were enriched in phospholipids, free sterols, and glycosphingolipids, but not in other neutral lipids or ceramides. In summary, whereas some of the enzymes in LB could participate in the metabolism of LB lipid precursors to hydrophobic barrier constituents, other may attack intercellular constituents, ultimately resulting in desquamation. The lipid profile of these organelles suggests that they deliver precursors of permeability barrier lipids to intercellular domains.

AB - Several problems have frustrated the isolation of lamellar bodies (LB) from mammalian epidermis. We obtained pellets enriched in intact LB by utilizing the staphylococcal epidermolytic toxin to provide intact, outer epidermal sheets, by controlled homogenization in a cell disrupter, and by passage of homogenates through a graded series of nuclepore filters (Science 221:962, 1983). Such preparations contained more intact LB than did fractions prepared by a variety of differential or sucrose/metrizamide discontinuous centrifugation methods. Initial characterization of the enzymatic content of this fraction revealed it to be enriched in certain hydrolytic enzymes (acid phosphatase, carboxypeptidase, cathepsin B, acid lipase, sphingomyelinase, and phospholipase A), but strikingly depleted in all sulfatases, β-glucuronidase, and the non-lysosomal protease, plasminogen activator. Thus, LB show some properties of lysosomes, although certain characteristic lysosomal enzymes are strikingly absent. Lamellar body fractions contained 2-3 times more lipid per unit weight than did homogenates, and were enriched in phospholipids, free sterols, and glycosphingolipids, but not in other neutral lipids or ceramides. In summary, whereas some of the enzymes in LB could participate in the metabolism of LB lipid precursors to hydrophobic barrier constituents, other may attack intercellular constituents, ultimately resulting in desquamation. The lipid profile of these organelles suggests that they deliver precursors of permeability barrier lipids to intercellular domains.

UR - http://www.scopus.com/inward/record.url?scp=0021784419&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021784419&partnerID=8YFLogxK

M3 - Article

C2 - 4045217

AN - SCOPUS:0021784419

VL - 85

SP - 289

EP - 294

JO - Journal of Investigative Dermatology

JF - Journal of Investigative Dermatology

SN - 0022-202X

IS - 4

ER -