Structure of Human Apolipoprotein D: Locations of the Intermolecular and Intramolecular Disulfide Links

Chao Yuh Yang; Gu Zi-Wei; Francisco Blanco-Vaca; Simon J. Gaskell; Manlan Yang; John B. Massey; Antonio Gotto; Henry J. Pownall

doi:10.1021/bi00207a011

Structure of Human Apolipoprotein D: Locations of the Intermolecular and Intramolecular Disulfide Links

Chao Yuh Yang, Gu Zi-Wei, Francisco Blanco-Vaca, Simon J. Gaskell, Manlan Yang, John B. Massey, Antonio Gotto, Henry J. Pownall

Research output: Contribution to journal › Article › peer-review

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Abstract

We have determined the primary structure of human apolipoprotein D (apoD) by aligning peptides derived from digestions by cyanogen bromide, trypsin, and chymotrypsin. Our results confirm the primary structure derived from cDNA [Drayna et al. (1986) J. Biol. Chem. 261, 16535-16539]. ApoD consists of 169 amino acid residues, including 5 cysteines. Tryptic peptide analysis indicated that Cys41 and Cys16 are joined by a disulfide bridge. Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. In addition, sites of N-glycosylation were found at Asn45 and Asn78. Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt α-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. We conclude that apoD binds to lipoprotein surfaces through structures other than α-helices, such as disulfide links.

Original language	English (US)
Pages (from-to)	12451-12455
Number of pages	5
Journal	Biochemistry
Volume	33
Issue number	41
DOIs	https://doi.org/10.1021/bi00207a011
State	Published - Oct 1 1994

ASJC Scopus subject areas

Biochemistry

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title = "Structure of Human Apolipoprotein D: Locations of the Intermolecular and Intramolecular Disulfide Links",

abstract = "We have determined the primary structure of human apolipoprotein D (apoD) by aligning peptides derived from digestions by cyanogen bromide, trypsin, and chymotrypsin. Our results confirm the primary structure derived from cDNA [Drayna et al. (1986) J. Biol. Chem. 261, 16535-16539]. ApoD consists of 169 amino acid residues, including 5 cysteines. Tryptic peptide analysis indicated that Cys41 and Cys16 are joined by a disulfide bridge. Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. In addition, sites of N-glycosylation were found at Asn45 and Asn78. Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt α-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. We conclude that apoD binds to lipoprotein surfaces through structures other than α-helices, such as disulfide links.",

author = "Yang, {Chao Yuh} and Gu Zi-Wei and Francisco Blanco-Vaca and Gaskell, {Simon J.} and Manlan Yang and Massey, {John B.} and Antonio Gotto and Pownall, {Henry J.}",

year = "1994",

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T2 - Locations of the Intermolecular and Intramolecular Disulfide Links

AU - Yang, Chao Yuh

AU - Zi-Wei, Gu

AU - Blanco-Vaca, Francisco

AU - Gaskell, Simon J.

AU - Yang, Manlan

AU - Massey, John B.

AU - Gotto, Antonio

AU - Pownall, Henry J.

PY - 1994/10/1

Y1 - 1994/10/1

N2 - We have determined the primary structure of human apolipoprotein D (apoD) by aligning peptides derived from digestions by cyanogen bromide, trypsin, and chymotrypsin. Our results confirm the primary structure derived from cDNA [Drayna et al. (1986) J. Biol. Chem. 261, 16535-16539]. ApoD consists of 169 amino acid residues, including 5 cysteines. Tryptic peptide analysis indicated that Cys41 and Cys16 are joined by a disulfide bridge. Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. In addition, sites of N-glycosylation were found at Asn45 and Asn78. Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt α-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. We conclude that apoD binds to lipoprotein surfaces through structures other than α-helices, such as disulfide links.

AB - We have determined the primary structure of human apolipoprotein D (apoD) by aligning peptides derived from digestions by cyanogen bromide, trypsin, and chymotrypsin. Our results confirm the primary structure derived from cDNA [Drayna et al. (1986) J. Biol. Chem. 261, 16535-16539]. ApoD consists of 169 amino acid residues, including 5 cysteines. Tryptic peptide analysis indicated that Cys41 and Cys16 are joined by a disulfide bridge. Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. In addition, sites of N-glycosylation were found at Asn45 and Asn78. Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt α-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. We conclude that apoD binds to lipoprotein surfaces through structures other than α-helices, such as disulfide links.

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Structure of Human Apolipoprotein D: Locations of the Intermolecular and Intramolecular Disulfide Links

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