Newly synthesized calsequestrin, destined for the sarcoplasmic reticulum, is contained in early/intermediate Golgi-derived clathrin-coated vesicles

K. Thomas, Javier Navarro, R. J J Benson, K. P. Campbell, R. L. Rotundo, R. E. Fine

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

We have examined the possible role of clathrin-coated vesicles (CVs) in the genesis of the sarcoplasmic reticulum (SR) in developing chick skeletal myotubes. Calsequestrin (CSQ) a luminal Ca2+ binding protein of the terminal SR cisternae, is contained within the vesicle lumen of skeletal muscle CVs in substantial amounts, approximately four molecules/CV. Employing 3-day cultures of chick skeletal myotubes we demonstrate that after a 30-min labeling with [35S]methionine and cysteine, radioactivity in CSQ remains high in the CVs 45 min later and then declines, while labeled CSQ in the SR continues to rise. No CSQ appears to be secreted. All of the CSQ in both the CVs and SR is sensitive to the activity of endoglycosidase H, and a significant fraction also binds to wheat germ agglutinin. Based on these results, we discuss the hypothesis that a selective CV-mediated pathway exists in developing skeletal muscle cells for the transport of CSQ from the early/intermediate Golgi apparatus to the SR.

Original languageEnglish (US)
Pages (from-to)3140-3145
Number of pages6
JournalJournal of Biological Chemistry
Volume264
Issue number6
StatePublished - 1989
Externally publishedYes

Fingerprint

Calsequestrin
Clathrin-Coated Vesicles
Clathrin
Coated Vesicles
Sarcoplasmic Reticulum
Skeletal Muscle Fibers
Muscle
Skeletal Muscle
Wheat Germ Agglutinins
Glycoside Hydrolases
Radioactivity
Golgi Apparatus
Methionine
Muscle Cells
Labeling
Cysteine
Carrier Proteins
Cells
Molecules

ASJC Scopus subject areas

  • Biochemistry

Cite this

Newly synthesized calsequestrin, destined for the sarcoplasmic reticulum, is contained in early/intermediate Golgi-derived clathrin-coated vesicles. / Thomas, K.; Navarro, Javier; Benson, R. J J; Campbell, K. P.; Rotundo, R. L.; Fine, R. E.

In: Journal of Biological Chemistry, Vol. 264, No. 6, 1989, p. 3140-3145.

Research output: Contribution to journalArticle

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AU - Navarro, Javier

AU - Benson, R. J J

AU - Campbell, K. P.

AU - Rotundo, R. L.

AU - Fine, R. E.

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N2 - We have examined the possible role of clathrin-coated vesicles (CVs) in the genesis of the sarcoplasmic reticulum (SR) in developing chick skeletal myotubes. Calsequestrin (CSQ) a luminal Ca2+ binding protein of the terminal SR cisternae, is contained within the vesicle lumen of skeletal muscle CVs in substantial amounts, approximately four molecules/CV. Employing 3-day cultures of chick skeletal myotubes we demonstrate that after a 30-min labeling with [35S]methionine and cysteine, radioactivity in CSQ remains high in the CVs 45 min later and then declines, while labeled CSQ in the SR continues to rise. No CSQ appears to be secreted. All of the CSQ in both the CVs and SR is sensitive to the activity of endoglycosidase H, and a significant fraction also binds to wheat germ agglutinin. Based on these results, we discuss the hypothesis that a selective CV-mediated pathway exists in developing skeletal muscle cells for the transport of CSQ from the early/intermediate Golgi apparatus to the SR.

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