Partial characterization of a vicilin-like glycoprotein from seeds of flowering tobacco (Nicotiana sylvestris)

Jared Q. Gerlach; Veer P. Bhavanandan; Paul A. Haynes; Lokesh Joshi

doi:10.1155/2009/560394

Partial characterization of a vicilin-like glycoprotein from seeds of flowering tobacco (Nicotiana sylvestris)

Jared Q. Gerlach, Veer P. Bhavanandan, Paul A. Haynes, Lokesh Joshi

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Abstract

A vicilin-like glycoprotein from the seeds of Nicotiana sylvestris, flowering tobacco, has been identified using nanoLC/ESI-MS/MS. Sequences from a fragment of protein demonstrated homology with vicilins from other members of the Solanaceae family, notably potato (Solanum demissum). Reducing and nonreducing SDS-PAGE analyses of the identified protein indicated that fragments resulting from in situ proteolytic processing are joined by intrachain disulphide bonds. Staining with Con A lectin was specifically inhibited by mannose suggested the presence of N-linked glycosylation which was confirmed by carbohydrate compositional analysis of PVDF-bound protein subunits. HPAEC-PAD analysis of the monosaccharides released from the glycoprotein by acid hydrolysis revealed glucosamine and mannose. N-acetylglucosamine termination of attached oligosaccharides was further verified by inhibitable WGA lectin staining. Immunostaining of PVDF-bound N. sylvestris proteins with antibodies against G. max total protein demonstrated cross-staining at masses corresponding to fragments from the proteolytically processed protein subunits.

Original language	English
Article number	560394
Pages (from-to)	1-12
Number of pages	12
Journal	Journal of Botany
Volume	2009
DOIs	https://doi.org/10.1155/2009/560394
Publication status	Published - 2009

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Copyright the Author(s) 2009. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

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10.1155/2009/560394

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@article{ca4385205efa4fb988e27b693db7d5c5,

title = "Partial characterization of a vicilin-like glycoprotein from seeds of flowering tobacco (Nicotiana sylvestris)",

abstract = "A vicilin-like glycoprotein from the seeds of Nicotiana sylvestris, flowering tobacco, has been identified using nanoLC/ESI-MS/MS. Sequences from a fragment of protein demonstrated homology with vicilins from other members of the Solanaceae family, notably potato (Solanum demissum). Reducing and nonreducing SDS-PAGE analyses of the identified protein indicated that fragments resulting from in situ proteolytic processing are joined by intrachain disulphide bonds. Staining with Con A lectin was specifically inhibited by mannose suggested the presence of N-linked glycosylation which was confirmed by carbohydrate compositional analysis of PVDF-bound protein subunits. HPAEC-PAD analysis of the monosaccharides released from the glycoprotein by acid hydrolysis revealed glucosamine and mannose. N-acetylglucosamine termination of attached oligosaccharides was further verified by inhibitable WGA lectin staining. Immunostaining of PVDF-bound N. sylvestris proteins with antibodies against G. max total protein demonstrated cross-staining at masses corresponding to fragments from the proteolytically processed protein subunits.",

author = "Gerlach, {Jared Q.} and Bhavanandan, {Veer P.} and Haynes, {Paul A.} and Lokesh Joshi",

note = "Copyright the Author(s) 2009. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.",

year = "2009",

doi = "10.1155/2009/560394",

language = "English",

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T1 - Partial characterization of a vicilin-like glycoprotein from seeds of flowering tobacco (Nicotiana sylvestris)

AU - Gerlach, Jared Q.

AU - Bhavanandan, Veer P.

AU - Haynes, Paul A.

AU - Joshi, Lokesh

N1 - Copyright the Author(s) 2009. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

PY - 2009

Y1 - 2009

N2 - A vicilin-like glycoprotein from the seeds of Nicotiana sylvestris, flowering tobacco, has been identified using nanoLC/ESI-MS/MS. Sequences from a fragment of protein demonstrated homology with vicilins from other members of the Solanaceae family, notably potato (Solanum demissum). Reducing and nonreducing SDS-PAGE analyses of the identified protein indicated that fragments resulting from in situ proteolytic processing are joined by intrachain disulphide bonds. Staining with Con A lectin was specifically inhibited by mannose suggested the presence of N-linked glycosylation which was confirmed by carbohydrate compositional analysis of PVDF-bound protein subunits. HPAEC-PAD analysis of the monosaccharides released from the glycoprotein by acid hydrolysis revealed glucosamine and mannose. N-acetylglucosamine termination of attached oligosaccharides was further verified by inhibitable WGA lectin staining. Immunostaining of PVDF-bound N. sylvestris proteins with antibodies against G. max total protein demonstrated cross-staining at masses corresponding to fragments from the proteolytically processed protein subunits.

AB - A vicilin-like glycoprotein from the seeds of Nicotiana sylvestris, flowering tobacco, has been identified using nanoLC/ESI-MS/MS. Sequences from a fragment of protein demonstrated homology with vicilins from other members of the Solanaceae family, notably potato (Solanum demissum). Reducing and nonreducing SDS-PAGE analyses of the identified protein indicated that fragments resulting from in situ proteolytic processing are joined by intrachain disulphide bonds. Staining with Con A lectin was specifically inhibited by mannose suggested the presence of N-linked glycosylation which was confirmed by carbohydrate compositional analysis of PVDF-bound protein subunits. HPAEC-PAD analysis of the monosaccharides released from the glycoprotein by acid hydrolysis revealed glucosamine and mannose. N-acetylglucosamine termination of attached oligosaccharides was further verified by inhibitable WGA lectin staining. Immunostaining of PVDF-bound N. sylvestris proteins with antibodies against G. max total protein demonstrated cross-staining at masses corresponding to fragments from the proteolytically processed protein subunits.

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DO - 10.1155/2009/560394

M3 - Article

SN - 2090-0120

VL - 2009

SP - 1

EP - 12

JO - Journal of Botany

JF - Journal of Botany

M1 - 560394

ER -

Partial characterization of a vicilin-like glycoprotein from seeds of flowering tobacco (Nicotiana sylvestris)

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