Identification and molecular characterization of a novel protein Saglin as a target of monoclonal antibodies affecting salivary gland infectivity of Plasmodium sporozoites

M. A. Okulate, D. E. Kalume, R. Reddy, T. Kristiansen, M. Bhattacharyya, R. Chaerkady, Akhilesh Pandey, N. Kumar

Research output: Contribution to journalArticle

Abstract

Molecular mechanisms underlying the interaction between malarial sporozoites and putative receptor(s) on the salivary glands of Anopheles gambiae remain largely unknown. In previous studies, a salivary gland protein of ∼100 kDa was identified as a putative target based on recognition of the protein by a monoclonal antibody (mAb) 2A3 that caused a ≥ 70% reduction in the average number of sporozoites per infected salivary gland when fed to mosquitoes. Using affinity purification we purified the target of this mAb from extracts of female A. gambiae salivary glands and it was found to be a novel protein by tandem mass spectrometric analysis. Biochemical and molecular characterization of the 100 kDa protein showed that this molecule, designated Saglin, exists as a disulphide-bonded homodimer of 50 kDa subunits. The ability to form homodimers was retained even in the recombinant Saglin expressed in mammalian cells (HEK293). The amino acid sequence of Saglin contains a signal peptide suggesting that Saglin is a secreted protein. If Saglin is indeed involved in the process of invasion of A. gambiae salivary glands by sporozoites of Plasmodium, it could provide a novel target for future investigations aimed at interruption of malaria transmission.

Original languageEnglish (US)
Pages (from-to)711-722
Number of pages12
JournalInsect Molecular Biology
Volume16
Issue number6
DOIs
StatePublished - Dec 2007

Fingerprint

Sporozoites
Plasmodium
sporozoites
salivary glands
Salivary Glands
Anopheles gambiae
monoclonal antibodies
pathogenicity
Monoclonal Antibodies
Proteins
proteins
Salivary Proteins and Peptides
HEK293 Cells
Protein Sorting Signals
Culicidae
Disulfides
Malaria
Purification
Amino Acid Sequence
signal peptide

Keywords

  • Anopheline
  • Malaria
  • Receptor
  • Salivary gland
  • Sporozoite

ASJC Scopus subject areas

  • Insect Science
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Identification and molecular characterization of a novel protein Saglin as a target of monoclonal antibodies affecting salivary gland infectivity of Plasmodium sporozoites. / Okulate, M. A.; Kalume, D. E.; Reddy, R.; Kristiansen, T.; Bhattacharyya, M.; Chaerkady, R.; Pandey, Akhilesh; Kumar, N.

In: Insect Molecular Biology, Vol. 16, No. 6, 12.2007, p. 711-722.

Research output: Contribution to journalArticle

Okulate, M. A. ; Kalume, D. E. ; Reddy, R. ; Kristiansen, T. ; Bhattacharyya, M. ; Chaerkady, R. ; Pandey, Akhilesh ; Kumar, N. / Identification and molecular characterization of a novel protein Saglin as a target of monoclonal antibodies affecting salivary gland infectivity of Plasmodium sporozoites. In: Insect Molecular Biology. 2007 ; Vol. 16, No. 6. pp. 711-722.
@article{2a0b4de0992d4a14b0e91d6795f116b7,
title = "Identification and molecular characterization of a novel protein Saglin as a target of monoclonal antibodies affecting salivary gland infectivity of Plasmodium sporozoites",
abstract = "Molecular mechanisms underlying the interaction between malarial sporozoites and putative receptor(s) on the salivary glands of Anopheles gambiae remain largely unknown. In previous studies, a salivary gland protein of ∼100 kDa was identified as a putative target based on recognition of the protein by a monoclonal antibody (mAb) 2A3 that caused a ≥ 70{\%} reduction in the average number of sporozoites per infected salivary gland when fed to mosquitoes. Using affinity purification we purified the target of this mAb from extracts of female A. gambiae salivary glands and it was found to be a novel protein by tandem mass spectrometric analysis. Biochemical and molecular characterization of the 100 kDa protein showed that this molecule, designated Saglin, exists as a disulphide-bonded homodimer of 50 kDa subunits. The ability to form homodimers was retained even in the recombinant Saglin expressed in mammalian cells (HEK293). The amino acid sequence of Saglin contains a signal peptide suggesting that Saglin is a secreted protein. If Saglin is indeed involved in the process of invasion of A. gambiae salivary glands by sporozoites of Plasmodium, it could provide a novel target for future investigations aimed at interruption of malaria transmission.",
keywords = "Anopheline, Malaria, Receptor, Salivary gland, Sporozoite",
author = "Okulate, {M. A.} and Kalume, {D. E.} and R. Reddy and T. Kristiansen and M. Bhattacharyya and R. Chaerkady and Akhilesh Pandey and N. Kumar",
year = "2007",
month = "12",
doi = "10.1111/j.1365-2583.2007.00765.x",
language = "English (US)",
volume = "16",
pages = "711--722",
journal = "Insect Molecular Biology",
issn = "0962-1075",
publisher = "Wiley-Blackwell",
number = "6",

}

TY - JOUR

T1 - Identification and molecular characterization of a novel protein Saglin as a target of monoclonal antibodies affecting salivary gland infectivity of Plasmodium sporozoites

AU - Okulate, M. A.

AU - Kalume, D. E.

AU - Reddy, R.

AU - Kristiansen, T.

AU - Bhattacharyya, M.

AU - Chaerkady, R.

AU - Pandey, Akhilesh

AU - Kumar, N.

PY - 2007/12

Y1 - 2007/12

N2 - Molecular mechanisms underlying the interaction between malarial sporozoites and putative receptor(s) on the salivary glands of Anopheles gambiae remain largely unknown. In previous studies, a salivary gland protein of ∼100 kDa was identified as a putative target based on recognition of the protein by a monoclonal antibody (mAb) 2A3 that caused a ≥ 70% reduction in the average number of sporozoites per infected salivary gland when fed to mosquitoes. Using affinity purification we purified the target of this mAb from extracts of female A. gambiae salivary glands and it was found to be a novel protein by tandem mass spectrometric analysis. Biochemical and molecular characterization of the 100 kDa protein showed that this molecule, designated Saglin, exists as a disulphide-bonded homodimer of 50 kDa subunits. The ability to form homodimers was retained even in the recombinant Saglin expressed in mammalian cells (HEK293). The amino acid sequence of Saglin contains a signal peptide suggesting that Saglin is a secreted protein. If Saglin is indeed involved in the process of invasion of A. gambiae salivary glands by sporozoites of Plasmodium, it could provide a novel target for future investigations aimed at interruption of malaria transmission.

AB - Molecular mechanisms underlying the interaction between malarial sporozoites and putative receptor(s) on the salivary glands of Anopheles gambiae remain largely unknown. In previous studies, a salivary gland protein of ∼100 kDa was identified as a putative target based on recognition of the protein by a monoclonal antibody (mAb) 2A3 that caused a ≥ 70% reduction in the average number of sporozoites per infected salivary gland when fed to mosquitoes. Using affinity purification we purified the target of this mAb from extracts of female A. gambiae salivary glands and it was found to be a novel protein by tandem mass spectrometric analysis. Biochemical and molecular characterization of the 100 kDa protein showed that this molecule, designated Saglin, exists as a disulphide-bonded homodimer of 50 kDa subunits. The ability to form homodimers was retained even in the recombinant Saglin expressed in mammalian cells (HEK293). The amino acid sequence of Saglin contains a signal peptide suggesting that Saglin is a secreted protein. If Saglin is indeed involved in the process of invasion of A. gambiae salivary glands by sporozoites of Plasmodium, it could provide a novel target for future investigations aimed at interruption of malaria transmission.

KW - Anopheline

KW - Malaria

KW - Receptor

KW - Salivary gland

KW - Sporozoite

UR - http://www.scopus.com/inward/record.url?scp=37349042224&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=37349042224&partnerID=8YFLogxK

U2 - 10.1111/j.1365-2583.2007.00765.x

DO - 10.1111/j.1365-2583.2007.00765.x

M3 - Article

C2 - 18093000

AN - SCOPUS:37349042224

VL - 16

SP - 711

EP - 722

JO - Insect Molecular Biology

JF - Insect Molecular Biology

SN - 0962-1075

IS - 6

ER -