Variation in the glycosylation of the murine sex-limited protein: Comparison with the fourth component of murine complement

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Abstract

The murine sex-limited protein (SIp) is a hemolytically nonfunctional homologue of the fourth component of complement (C4). Two cogenic mouse strains, B10.BUA1 (H-2(w16)) and B10.KPB128 (H-2(w19)), which have been previously shown to share a variant form of C4 (Karp et al., J. Biol. Chem., 257: 7330-7335), were examined and found to also produce a variant form of SIp. SIp molecules isolated from the plasma or peritoneal macrophage cultures from these strains have an α-chain ~ 2,000 daltons smaller than th α-chain of SIp from H-2(d) or H-2(w7) mice as judged by sodium dodecyl sulfate (NaDodSO4)/polyacrylamide gel electrophoresis. Expression of this SIp was constitutive, i.e., not regulated by androgen, and is cis-dominant in F1 hybrid mice. Autolysis of the different relative molecular mass (M(r)) α-chains at the internal thiolester produced similar M(r) amino-terminal fragments and different M(r) carboxy-terminal fragments. Deglycosylation of the α-chains with trifluoromethanesulfonic acid eliminated most, if not all, the M(r) difference. The M(r) difference was also manifested by the intracellular precursors of SIp and could be eliminated by endoglycosidase H (endo H) treatment. The number of oligosaccharides on the SIp α-chain was deduced by limited endo H treatment of SIp synthesized in the presence of swainsonine, a plant alkaloid that prevents maturation of complex-type oligosaccharides. This method is a simple way to enumerate the complex-type, N-linked oligosaccharides on glycoproteins. The genetic variation in the glycosylation of C4, and a scheme depicting some of the structural differences among these molecules was developed.

Original languageEnglish (US)
Pages (from-to)1405-1410
Number of pages6
JournalJournal of Immunology
Volume131
Issue number3
StatePublished - 1983

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Glycosylation
Proteins
Oligosaccharides
Glycoside Hydrolases
Swainsonine
Complement C4
Autolysis
Peritoneal Macrophages
Alkaloids
Sodium Dodecyl Sulfate
Androgens
Polyacrylamide Gel Electrophoresis
Glycoproteins

ASJC Scopus subject areas

  • Immunology

Cite this

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title = "Variation in the glycosylation of the murine sex-limited protein: Comparison with the fourth component of murine complement",
abstract = "The murine sex-limited protein (SIp) is a hemolytically nonfunctional homologue of the fourth component of complement (C4). Two cogenic mouse strains, B10.BUA1 (H-2(w16)) and B10.KPB128 (H-2(w19)), which have been previously shown to share a variant form of C4 (Karp et al., J. Biol. Chem., 257: 7330-7335), were examined and found to also produce a variant form of SIp. SIp molecules isolated from the plasma or peritoneal macrophage cultures from these strains have an α-chain ~ 2,000 daltons smaller than th α-chain of SIp from H-2(d) or H-2(w7) mice as judged by sodium dodecyl sulfate (NaDodSO4)/polyacrylamide gel electrophoresis. Expression of this SIp was constitutive, i.e., not regulated by androgen, and is cis-dominant in F1 hybrid mice. Autolysis of the different relative molecular mass (M(r)) α-chains at the internal thiolester produced similar M(r) amino-terminal fragments and different M(r) carboxy-terminal fragments. Deglycosylation of the α-chains with trifluoromethanesulfonic acid eliminated most, if not all, the M(r) difference. The M(r) difference was also manifested by the intracellular precursors of SIp and could be eliminated by endoglycosidase H (endo H) treatment. The number of oligosaccharides on the SIp α-chain was deduced by limited endo H treatment of SIp synthesized in the presence of swainsonine, a plant alkaloid that prevents maturation of complex-type oligosaccharides. This method is a simple way to enumerate the complex-type, N-linked oligosaccharides on glycoproteins. The genetic variation in the glycosylation of C4, and a scheme depicting some of the structural differences among these molecules was developed.",
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T1 - Variation in the glycosylation of the murine sex-limited protein

T2 - Comparison with the fourth component of murine complement

AU - Karp, D. R.

PY - 1983

Y1 - 1983

N2 - The murine sex-limited protein (SIp) is a hemolytically nonfunctional homologue of the fourth component of complement (C4). Two cogenic mouse strains, B10.BUA1 (H-2(w16)) and B10.KPB128 (H-2(w19)), which have been previously shown to share a variant form of C4 (Karp et al., J. Biol. Chem., 257: 7330-7335), were examined and found to also produce a variant form of SIp. SIp molecules isolated from the plasma or peritoneal macrophage cultures from these strains have an α-chain ~ 2,000 daltons smaller than th α-chain of SIp from H-2(d) or H-2(w7) mice as judged by sodium dodecyl sulfate (NaDodSO4)/polyacrylamide gel electrophoresis. Expression of this SIp was constitutive, i.e., not regulated by androgen, and is cis-dominant in F1 hybrid mice. Autolysis of the different relative molecular mass (M(r)) α-chains at the internal thiolester produced similar M(r) amino-terminal fragments and different M(r) carboxy-terminal fragments. Deglycosylation of the α-chains with trifluoromethanesulfonic acid eliminated most, if not all, the M(r) difference. The M(r) difference was also manifested by the intracellular precursors of SIp and could be eliminated by endoglycosidase H (endo H) treatment. The number of oligosaccharides on the SIp α-chain was deduced by limited endo H treatment of SIp synthesized in the presence of swainsonine, a plant alkaloid that prevents maturation of complex-type oligosaccharides. This method is a simple way to enumerate the complex-type, N-linked oligosaccharides on glycoproteins. The genetic variation in the glycosylation of C4, and a scheme depicting some of the structural differences among these molecules was developed.

AB - The murine sex-limited protein (SIp) is a hemolytically nonfunctional homologue of the fourth component of complement (C4). Two cogenic mouse strains, B10.BUA1 (H-2(w16)) and B10.KPB128 (H-2(w19)), which have been previously shown to share a variant form of C4 (Karp et al., J. Biol. Chem., 257: 7330-7335), were examined and found to also produce a variant form of SIp. SIp molecules isolated from the plasma or peritoneal macrophage cultures from these strains have an α-chain ~ 2,000 daltons smaller than th α-chain of SIp from H-2(d) or H-2(w7) mice as judged by sodium dodecyl sulfate (NaDodSO4)/polyacrylamide gel electrophoresis. Expression of this SIp was constitutive, i.e., not regulated by androgen, and is cis-dominant in F1 hybrid mice. Autolysis of the different relative molecular mass (M(r)) α-chains at the internal thiolester produced similar M(r) amino-terminal fragments and different M(r) carboxy-terminal fragments. Deglycosylation of the α-chains with trifluoromethanesulfonic acid eliminated most, if not all, the M(r) difference. The M(r) difference was also manifested by the intracellular precursors of SIp and could be eliminated by endoglycosidase H (endo H) treatment. The number of oligosaccharides on the SIp α-chain was deduced by limited endo H treatment of SIp synthesized in the presence of swainsonine, a plant alkaloid that prevents maturation of complex-type oligosaccharides. This method is a simple way to enumerate the complex-type, N-linked oligosaccharides on glycoproteins. The genetic variation in the glycosylation of C4, and a scheme depicting some of the structural differences among these molecules was developed.

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