Salivary homogenates of the adult female mosquito Anopheles albimanus have been shown previously to contain a vasodilatory activity associated with a catechol oxidase/peroxidase activity. We have now purified the salivary peroxidase using high-performance liquid chromatography. The pure enzyme is able to relax rabbit aortic rings pre-constricted with norepinephrine. The peroxidase has a relative molecular mass of 66 907 as estimated by mass spectrometry. Amino-terminal sequencing allowed us to design oligonucleotide probes for isolation of cDNA clones derived from the salivary gland mRNA from female mosquitoes. The full sequence of the cDNA demonstrated homology between A. albimanus salivary peroxidase and several members of the myeloperoxidase gene family. A close comparison of A. albimanus salivary peroxidase with canine myeloperoxidase, for which the crystal structure is known, showed that all six disulfide bridges were conserved and demonstrated identity for all five residues associated with a Ca2+-binding site. In addition, 16 of 26 residues shown to be in close proximity to the heme moiety in the canine myeloperoxidase were identical. We conclude that the salivary peroxidase of A. albimanus belongs to the myeloperoxidase gene family. Other possible functions for this molecule in blood feeding are discussed.

Altschul
S. F.
,
Madden
T. L.
,
Schaffer
A. A.
,
Zhang
J.
,
Zhang
Z.
,
Miller
W.
,
Lipman
D. J.
(
1997
).
Gapped BLAST and PSI-BLAST: a new generation of protein database search programs
.
Nucleic Acids Res
25
,
3389
–.
Champagne
D.
,
Nussenzveig
R. H.
,
Ribeiro
J. M. C.
(
1995
).
Purification, characterization and cloning of nitric oxide-carrying heme proteins (nitrophorins) from salivary glands of the blood sucking insect Rhodnius prolixus
.
J. Biol. Chem
270
,
8691
–.
Champagne
D.
,
Ribeiro
J. M. C.
(
1994
).
Sialokinins I and II: Two salivary tachykinins from the Yellow Fever mosquito, Aedes aegypti
.
Proc. Natl. Acad. Sci. USA
91
,
138
–.
Cupp
M.
,
Ribeiro
J.
,
Champagne
D.
,
Cupp
E.
(
1998
).
Analyses of cDNA and recombinant protein for a potent vasoactive protein in saliva of a blood-feeding black fly, Simulium vittatum
.
J. Exp. Biol
201
,
1553
–.
Dickinson
R. G.
,
O'Hagan
J. E.
,
Shotz
M.
,
Binnington
K. C.
,
Hegarty
M. P.
(
1976
).
Prostaglandin in the saliva of the cattle tick Boophilus microplus
.
Aust. J. Exp. Biol. Med. Sci
54
,
475
–.
Henderson
W. R.
,
Jorg
A.
,
Klebanoff
S. J.
(
1982
).
Eosinophil peroxidase-mediated inactivation of leukotrienes B4, C4 and D4
.
J. Immunol
128
,
2609
–.
Henderson
W. R.
,
Klebanoff
S. J.
(
1983
).
Leukotriene production and inactivation by normal, chronic granulomatous disease and myeloperoxidase-deficient neutrophils
.
J. Biol. Chem
258
,
13522
–.
Higgs
G. A.
,
Vane
J. R.
,
Hart
R. J.
,
Porter
C.
,
Wilson
R. G.
(
1976
).
Prostaglandins in the saliva of the cattle tick, Boophilus microplus (Canestrini) (Acarina, Ixodidae)
.
Bull. Ent. Res
66
,
665
–.
Hsi
L. C.
,
Tsai
A. L.
,
Kulmacz
R. J.
,
English
D. G.
,
Siefker
A. O.
,
Otto
J. C.
,
Smith
W. L.
(
1993
).
Trp387 and the putative leucine zippers of PGH synthases-1 and-2
.
J. Lipid Mediat
6
,
131
–.
Johansson
M. W.
,
Lind
M. I.
,
Holmblad
T.
,
Thornqvist
P. O.
,
Soderhall
K.
(
1995
).
Peroxinectin, a novel cell adhesion816protein from crayfish blood
.
Biochem. Biophys. Res. Commun
216
,
1079
–.
Johansson
M. W.
,
Patarroyo
M.
,
Oberg
F.
,
Siegbahn
A.
,
Nilsson
K.
(
1997
).
Myeloperoxidase mediates cell adhesion via the alpha M beta 2 integrin (Mac-1, CD11b/CD18)
.
J. Cell Sci
110
,
1133
–.
Lerner
E. A.
,
Shoemaker
C. B.
(
1992
).
Maxadilan: Cloning and functional expression of the gene encoding this potent vasodilator peptide
.
J. Biol. Chem
267
,
1062
–.
Marinotti
O.
,
James
A.
,
Ribeiro
J. M. C.
(
1990
).
Diet and salivation in female Aedes aegypti mosquitoes
.
J. Insect Physiol
36
,
545
–.
Marshall
R. D.
(
1972
).
Glycoproteins
.
Annu. Rev. Biochem
41
,
673
–.
Moncada
S.
,
Palmer
R. M. J.
,
Higgs
E. A.
(
1991
).
Nitric oxide: Physiology, pathophysiology and pharmacology
.
Pharmac. Rev
43
,
109
–.
Ribeiro
J. M. C.
(
1987
).
Role of arthropod saliva in blood feeding
.
Annu. Rev. Ent
32
,
463
–.
Ribeiro
J. M. C.
(
1995
).
Blood-feeding arthropods: Live syringes or invertebrate pharmacologists?
.
Infect. Agents Dis
4
,
143
–.
Ribeiro
J. M. C.
,
Evans
P. M.
,
MacSwain
J. L.
,
Sauer
J.
(
1992
).
Amblyomma americanum: Characterization of salivary prostaglandins E2 and F2alpha by RP-HPLC/bioassay and gas chromatography—mass spectrometry
.
Exp. Parasitol
74
,
112
–.
Ribeiro
J. M. C.
,
Hazzard
J. M. H.
,
Nussenzveig
R. H.
,
Champagne
D.
,
Walker
F. A.
(
1993
).
Reversible binding of nitric oxide by a salivary nitrosylhemeprotein from the blood sucking bug, Rhodnius prolixus
.
Science
260
,
539
–.
Ribeiro
J. M. C.
,
Makoul
G.
,
Robinson
D.
(
1988
).
Ixodesdammini: Evidence for salivary prostacyclin secretion
.
J. Parasitol
74
,
1068
–.
Ribeiro
J. M. C.
,
Nussenzveig
R. H.
(
1993
).
The salivary catechol oxidase/peroxidase activities of the mosquito, Anopheles albimanus
.
J. Exp. Biol
179
,
273
–.
Ribeiro
J. M. C.
,
Rossignol
P. A.
,
Spielman
A.
(
1985
).
Salivary gland apyrase determines probing time in anopheline mosquitoes
.
J. Insect Physiol
9
,
551
–.
Ribeiro
J. M. C.
,
Sarkis
J. J. F.
,
Rossignol
P. A.
,
Spielman
A.
(
1984
).
Salivary apyrase of Aedes aegypti: Characterization and secretory fate
.
Comp. Biochem. Physiol
79
,
81
–.
Valenzuela
J. G.
,
Ribeiro
J. M.
(
1998
).
Purification and cloning of the salivary nitrophorin from the hemipteran Cimex lectularius
.
J. Exp. Biol
201
,
2659
–.
Valenzuela
J.
,
Walker
F. A.
,
Ribeiro
J. M. C.
(
1995
).
The salivary vasodilator of the bed bug, Cimex lectularius
.
J. Exp. Biol
198
,
1519
–.
Walker
F. A.
,
Lo
M. W.
,
Ree
M. T.
(
1976
).
Electronic effects in transition metal porphyrins. The reactions of imidazoles and pyridines with a series of para-substituted tetraphenylporphyrin complexes of chloroiron(III)
.
J. Am. Chem. Soc
98
,
5552
–.
Webster
M.
,
Prado
E. S.
(
1970
).
Glandular kallikreins from horse and human urine and from hog pancreas
.
Meth. Enzymol
19
,
681
–.
Zeng
J.
,
Fenna
R. E.
(
1992
).
X-ray crystal structure of canine myeloperoxidase at 3Å resolution
.
J. Mol. Biol
226
,
185
–.
Zhang
S. D.
,
Kassis
J.
,
Olde
B.
,
Mellerick
D. M.
,
Odenwald
W. F.
(
1996
).
Pollux, a novel Drosophila adhesion molecule, belongs to a family of proteins expressed in plants, yeast, nematodes and mA
.
Genes Dev
10
,
1108
–.
This content is only available via PDF.