Identification of major outer surface proteins of Streptococcus agalactiae

MJG Hughes; JC Moore; JD Lane; R Wilson; PK Pribul; ZN Younes; RJ Dobson; P Everest; AJ Reason; JM Redfern; FM Greer; T Paxton; M Panico; HR Morris; RG Feldman; JD Santangelo

doi:10.1128/IAI.70.3.1254-1259.2002

Identification of major outer surface proteins of Streptococcus agalactiae

MJG Hughes, JC Moore, JD Lane, R Wilson, PK Pribul, ZN Younes, RJ Dobson, P Everest, AJ Reason, JM Redfern, FM Greer, T Paxton, M Panico, HR Morris, RG Feldman, JD Santangelo

Biostatistics & Health Informatics

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Abstract

To identify the major outer surface proteins of Streptococcus agalactiae (group B streptococcus), a proteomic analysis was undertaken. An extract of the outer surface proteins was separated by two-dimensional electrophoresis. The visualized spots were identified through a combination of peptide sequencing and reverse genetic methodologies. Of the 30 major spots identified as S. agalactiae specific, 27 have been identified. Six of these proteins, previously unidentified in S. agalactiae, were sequenced and cloned. These were ornithine carbamoyltransferase, phosphoglycerate kinase, nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase, purine nucleoside phosphorylase, enolase, and glucose-6-phosphate isomerase. Using a gram-positive expression system, we have overexpressed two of these proteins in an in vitro system. These recombinant, purified proteins were used to raise antisera. The identification of these proteins as residing on the outer surface was confirmed by the ability of the antisera to react against whole, live bacteria. Further, in a neonatal-animal model system, we demonstrate that some of these sera are protective against lethal doses of bacteria. These studies demonstrate the successful application of proteomics as a technique for identifying vaccine candidates.

Original language	English
Pages (from-to)	1254 - 1259
Number of pages	6
Journal	Infection and Immunity
Volume	70
Issue number	3
DOIs	https://doi.org/10.1128/IAI.70.3.1254-1259.2002
Publication status	Published - 2002

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Hughes, MJG., Moore, JC., Lane, JD., Wilson, R., Pribul, PK., Younes, ZN., Dobson, RJ., Everest, P., Reason, AJ., Redfern, JM., Greer, FM., Paxton, T., Panico, M., Morris, HR., Feldman, RG., & Santangelo, JD. (2002). Identification of major outer surface proteins of Streptococcus agalactiae. Infection and Immunity, 70(3), 1254 - 1259. https://doi.org/10.1128/IAI.70.3.1254-1259.2002

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title = "Identification of major outer surface proteins of Streptococcus agalactiae",

abstract = "To identify the major outer surface proteins of Streptococcus agalactiae (group B streptococcus), a proteomic analysis was undertaken. An extract of the outer surface proteins was separated by two-dimensional electrophoresis. The visualized spots were identified through a combination of peptide sequencing and reverse genetic methodologies. Of the 30 major spots identified as S. agalactiae specific, 27 have been identified. Six of these proteins, previously unidentified in S. agalactiae, were sequenced and cloned. These were ornithine carbamoyltransferase, phosphoglycerate kinase, nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase, purine nucleoside phosphorylase, enolase, and glucose-6-phosphate isomerase. Using a gram-positive expression system, we have overexpressed two of these proteins in an in vitro system. These recombinant, purified proteins were used to raise antisera. The identification of these proteins as residing on the outer surface was confirmed by the ability of the antisera to react against whole, live bacteria. Further, in a neonatal-animal model system, we demonstrate that some of these sera are protective against lethal doses of bacteria. These studies demonstrate the successful application of proteomics as a technique for identifying vaccine candidates.",

author = "MJG Hughes and JC Moore and JD Lane and R Wilson and PK Pribul and ZN Younes and RJ Dobson and P Everest and AJ Reason and JM Redfern and FM Greer and T Paxton and M Panico and HR Morris and RG Feldman and JD Santangelo",

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Hughes, MJG, Moore, JC, Lane, JD, Wilson, R, Pribul, PK, Younes, ZN, Dobson, RJ, Everest, P, Reason, AJ, Redfern, JM, Greer, FM, Paxton, T, Panico, M, Morris, HR, Feldman, RG & Santangelo, JD 2002, 'Identification of major outer surface proteins of Streptococcus agalactiae', Infection and Immunity, vol. 70, no. 3, pp. 1254 - 1259. https://doi.org/10.1128/IAI.70.3.1254-1259.2002

TY - JOUR

T1 - Identification of major outer surface proteins of Streptococcus agalactiae

AU - Hughes, MJG

AU - Moore, JC

AU - Lane, JD

AU - Wilson, R

AU - Pribul, PK

AU - Younes, ZN

AU - Dobson, RJ

AU - Everest, P

AU - Reason, AJ

AU - Redfern, JM

AU - Greer, FM

AU - Paxton, T

AU - Panico, M

AU - Morris, HR

AU - Feldman, RG

AU - Santangelo, JD

PY - 2002

Y1 - 2002

N2 - To identify the major outer surface proteins of Streptococcus agalactiae (group B streptococcus), a proteomic analysis was undertaken. An extract of the outer surface proteins was separated by two-dimensional electrophoresis. The visualized spots were identified through a combination of peptide sequencing and reverse genetic methodologies. Of the 30 major spots identified as S. agalactiae specific, 27 have been identified. Six of these proteins, previously unidentified in S. agalactiae, were sequenced and cloned. These were ornithine carbamoyltransferase, phosphoglycerate kinase, nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase, purine nucleoside phosphorylase, enolase, and glucose-6-phosphate isomerase. Using a gram-positive expression system, we have overexpressed two of these proteins in an in vitro system. These recombinant, purified proteins were used to raise antisera. The identification of these proteins as residing on the outer surface was confirmed by the ability of the antisera to react against whole, live bacteria. Further, in a neonatal-animal model system, we demonstrate that some of these sera are protective against lethal doses of bacteria. These studies demonstrate the successful application of proteomics as a technique for identifying vaccine candidates.

AB - To identify the major outer surface proteins of Streptococcus agalactiae (group B streptococcus), a proteomic analysis was undertaken. An extract of the outer surface proteins was separated by two-dimensional electrophoresis. The visualized spots were identified through a combination of peptide sequencing and reverse genetic methodologies. Of the 30 major spots identified as S. agalactiae specific, 27 have been identified. Six of these proteins, previously unidentified in S. agalactiae, were sequenced and cloned. These were ornithine carbamoyltransferase, phosphoglycerate kinase, nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase, purine nucleoside phosphorylase, enolase, and glucose-6-phosphate isomerase. Using a gram-positive expression system, we have overexpressed two of these proteins in an in vitro system. These recombinant, purified proteins were used to raise antisera. The identification of these proteins as residing on the outer surface was confirmed by the ability of the antisera to react against whole, live bacteria. Further, in a neonatal-animal model system, we demonstrate that some of these sera are protective against lethal doses of bacteria. These studies demonstrate the successful application of proteomics as a technique for identifying vaccine candidates.

U2 - 10.1128/IAI.70.3.1254-1259.2002

DO - 10.1128/IAI.70.3.1254-1259.2002

M3 - Article

VL - 70

SP - 1254

EP - 1259

JO - Infection and Immunity

JF - Infection and Immunity

IS - 3

ER -

Identification of major outer surface proteins of Streptococcus agalactiae

Abstract

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