Identification of specific Hep G2 cell binding regions in Plasmodium falciparum sporozoite-threonine-asparagine-rich protein (STARP).
It has been demonstrated that Plasmodium falciparum sporozoite threonine-asparagine-rich protein (PfSTARP) is located on the sporozoite surface. This protein's non-overlapping consecutive peptides were synthesised and tested in Hep G2 cell binding assays. Twelve high activity binding peptides (HABPs) were identified in the resulting 31 peptides. Three were found in 5' non-repeat region (amino acids 41-80). Peptides 20546 (41VIKHNRFLSEYQSNFLGGGY(60)), 20547 (61SAALKLVNSKKSGTNVNVTKY(80)) and 20548 (81NSENTNTNNNIPESSSTYTN(100)) were located in the conserved amino terminal region, as well as peptide 20548 which shared the sequence with the M region (amino acids 85-134). Six HABPs were located in region 10 (Rp10) (STDNNNTKTI). HABPs 20569 (501TSDDELNKDSCDYSEEKENI(520)) and 20570 (521KSMINAYLDKLDLETVRKIH(40)) were found in 3' non-repeat region. All these HABPs showed saturable binding and presented dissociation constants between 18 and 219 nM. The number of binding sites per Hep G2 cell ranged from 45000 to 370000. High binding peptides' critical amino acids involved in Hep G2 cell binding were determined by competition binding assays. SDS-PAGE results showed that both peptides 20570 and 20547 had at least two different sets of 44 and 38 kDa HABP receptors on Hep G2 cells. Specific modification of peptide 20546 and 20570 critical binding residues rendered these peptides immunogenic in Aotus monkeys, inducing high antibody titres against sporozoites, as assessed by IFA.
López
Ramsés
R
Fundación Instituto de Inmunología de Colombia (FIDIC), Universidad Nacional de Colombia, Carrera 50 No 26-00, Bogotá, Colombia.
Garcia
Javier
J
Puentes
Alvaro
A
Curtidor
Hernando
H
Ocampo
Marisol
M
Vera
Ricardo
R
Rodriguez
Luis Eduardo
LE
Suarez
Jorge
J
Urquiza
Mauricio
M
Rodríguez
Ana Liliana
AL
Reyes
Claudia Alexandra
CA
Granados
Carmen Giovana
CG
Patarroyo
Manuel E
ME
D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000276:immunology*; D001029:Aotus trivirgatus; D001665:Binding Sites; D005998:Glycine; D006515:Hepatitis B virus / Q000276:immunology; D006801:Humans; D017780:Malaria Vaccines / Q000494:pharmacology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D010963:Plasmodium falciparum / Q000737:chemistry / Q000276:immunology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured
D000953:Antigens, Protozoan; D017780:Malaria Vaccines; D010446:Peptide Fragments; D011994:Recombinant Proteins; C109185:STARP antigen, Plasmodium falciparum; D005998:Glycine