Synthesis, secretion, and subcellular localization of serglycin proteoglycan in human endothelial cells

Barbara P. Schick, Joel Gradowski, James D. San Antonio

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

The serglycin proteoglycan is best known as a hematopoietic cell granule proteoglycan. It has been found that serglycin is synthesized by endothelial cells, is localized to cytoplasmic vesicles, and is constitutively secreted. Serglycin messenger RNA in human umbilical vein endothelial cells (HUVECs) and cultured human aortic endothelial cells was detected by reverse transcription-polymerase chain reaction. 35 S-sulfate-labeled secreted and intracellular proteoglycans were analyzed. It was found that 85% of the proteoglycans synthesized during culture were secreted. A core protein of the appropriate size for serglycin was detected by analysis of the chondroitinase-digested 35 S-sulfate-labeled HUVEC proteoglycans. This was the major core protein of the secreted chondroitin sulfate proteoglycans. Recombinant serglycin core protein was used to generate an antibody in chickens. A core protein identified by Western blotting of chondroitinase digests of HUVEC proteoglycans corresponded to the major 35 S-sulfate-labeled core protein. Identical results were obtained with 2 hematopoietic cell lines. Cyto-immunofluorescence showed cytoplasmic vesicular and perinuclear labeling in hematopoietic cells and HUVECs. The serglycin-containing vesicles in HUVECs are distinct from the Weibel-Palade bodies, which contain von Wille-brand factor. Confocal microscopy showed that tissue plasminogen activator was distributed similarly to serglycin. Serglycin may be important for the function of these vesicles and, once secreted, for the modulation of the activity of their constituents.

Original languageEnglish (US)
Pages (from-to)449-458
Number of pages10
JournalBlood
Volume97
Issue number2
DOIs
StatePublished - Jan 15 2001

Fingerprint

Endothelial cells
Proteoglycans
Endothelial Cells
Human Umbilical Vein Endothelial Cells
Chondroitinases and Chondroitin Lyases
Sulfates
Proteins
Weibel-Palade Bodies
Cytoplasmic Vesicles
Viral Core Proteins
Chondroitin Sulfate Proteoglycans
serglycin
Confocal microscopy
Polymerase chain reaction
Tissue Plasminogen Activator
Transcription
Confocal Microscopy
Labeling
Reverse Transcription
Fluorescent Antibody Technique

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

Cite this

Synthesis, secretion, and subcellular localization of serglycin proteoglycan in human endothelial cells. / Schick, Barbara P.; Gradowski, Joel; San Antonio, James D.

In: Blood, Vol. 97, No. 2, 15.01.2001, p. 449-458.

Research output: Contribution to journalArticle

Schick, Barbara P. ; Gradowski, Joel ; San Antonio, James D. / Synthesis, secretion, and subcellular localization of serglycin proteoglycan in human endothelial cells. In: Blood. 2001 ; Vol. 97, No. 2. pp. 449-458.
@article{e3c3d70d42e84b8d8447294ab5a5de2f,
title = "Synthesis, secretion, and subcellular localization of serglycin proteoglycan in human endothelial cells",
abstract = "The serglycin proteoglycan is best known as a hematopoietic cell granule proteoglycan. It has been found that serglycin is synthesized by endothelial cells, is localized to cytoplasmic vesicles, and is constitutively secreted. Serglycin messenger RNA in human umbilical vein endothelial cells (HUVECs) and cultured human aortic endothelial cells was detected by reverse transcription-polymerase chain reaction. 35 S-sulfate-labeled secreted and intracellular proteoglycans were analyzed. It was found that 85{\%} of the proteoglycans synthesized during culture were secreted. A core protein of the appropriate size for serglycin was detected by analysis of the chondroitinase-digested 35 S-sulfate-labeled HUVEC proteoglycans. This was the major core protein of the secreted chondroitin sulfate proteoglycans. Recombinant serglycin core protein was used to generate an antibody in chickens. A core protein identified by Western blotting of chondroitinase digests of HUVEC proteoglycans corresponded to the major 35 S-sulfate-labeled core protein. Identical results were obtained with 2 hematopoietic cell lines. Cyto-immunofluorescence showed cytoplasmic vesicular and perinuclear labeling in hematopoietic cells and HUVECs. The serglycin-containing vesicles in HUVECs are distinct from the Weibel-Palade bodies, which contain von Wille-brand factor. Confocal microscopy showed that tissue plasminogen activator was distributed similarly to serglycin. Serglycin may be important for the function of these vesicles and, once secreted, for the modulation of the activity of their constituents.",
author = "Schick, {Barbara P.} and Joel Gradowski and {San Antonio}, {James D.}",
year = "2001",
month = "1",
day = "15",
doi = "10.1182/blood.V97.2.449",
language = "English (US)",
volume = "97",
pages = "449--458",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "2",

}

TY - JOUR

T1 - Synthesis, secretion, and subcellular localization of serglycin proteoglycan in human endothelial cells

AU - Schick, Barbara P.

AU - Gradowski, Joel

AU - San Antonio, James D.

PY - 2001/1/15

Y1 - 2001/1/15

N2 - The serglycin proteoglycan is best known as a hematopoietic cell granule proteoglycan. It has been found that serglycin is synthesized by endothelial cells, is localized to cytoplasmic vesicles, and is constitutively secreted. Serglycin messenger RNA in human umbilical vein endothelial cells (HUVECs) and cultured human aortic endothelial cells was detected by reverse transcription-polymerase chain reaction. 35 S-sulfate-labeled secreted and intracellular proteoglycans were analyzed. It was found that 85% of the proteoglycans synthesized during culture were secreted. A core protein of the appropriate size for serglycin was detected by analysis of the chondroitinase-digested 35 S-sulfate-labeled HUVEC proteoglycans. This was the major core protein of the secreted chondroitin sulfate proteoglycans. Recombinant serglycin core protein was used to generate an antibody in chickens. A core protein identified by Western blotting of chondroitinase digests of HUVEC proteoglycans corresponded to the major 35 S-sulfate-labeled core protein. Identical results were obtained with 2 hematopoietic cell lines. Cyto-immunofluorescence showed cytoplasmic vesicular and perinuclear labeling in hematopoietic cells and HUVECs. The serglycin-containing vesicles in HUVECs are distinct from the Weibel-Palade bodies, which contain von Wille-brand factor. Confocal microscopy showed that tissue plasminogen activator was distributed similarly to serglycin. Serglycin may be important for the function of these vesicles and, once secreted, for the modulation of the activity of their constituents.

AB - The serglycin proteoglycan is best known as a hematopoietic cell granule proteoglycan. It has been found that serglycin is synthesized by endothelial cells, is localized to cytoplasmic vesicles, and is constitutively secreted. Serglycin messenger RNA in human umbilical vein endothelial cells (HUVECs) and cultured human aortic endothelial cells was detected by reverse transcription-polymerase chain reaction. 35 S-sulfate-labeled secreted and intracellular proteoglycans were analyzed. It was found that 85% of the proteoglycans synthesized during culture were secreted. A core protein of the appropriate size for serglycin was detected by analysis of the chondroitinase-digested 35 S-sulfate-labeled HUVEC proteoglycans. This was the major core protein of the secreted chondroitin sulfate proteoglycans. Recombinant serglycin core protein was used to generate an antibody in chickens. A core protein identified by Western blotting of chondroitinase digests of HUVEC proteoglycans corresponded to the major 35 S-sulfate-labeled core protein. Identical results were obtained with 2 hematopoietic cell lines. Cyto-immunofluorescence showed cytoplasmic vesicular and perinuclear labeling in hematopoietic cells and HUVECs. The serglycin-containing vesicles in HUVECs are distinct from the Weibel-Palade bodies, which contain von Wille-brand factor. Confocal microscopy showed that tissue plasminogen activator was distributed similarly to serglycin. Serglycin may be important for the function of these vesicles and, once secreted, for the modulation of the activity of their constituents.

UR - http://www.scopus.com/inward/record.url?scp=0035863954&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035863954&partnerID=8YFLogxK

U2 - 10.1182/blood.V97.2.449

DO - 10.1182/blood.V97.2.449

M3 - Article

VL - 97

SP - 449

EP - 458

JO - Blood

JF - Blood

SN - 0006-4971

IS - 2

ER -