PMID- 11329465
OWN - NLM
STAT- MEDLINE
DCOM- 20010719
LR  - 20181113
IS  - 1071-412X (Print)
IS  - 1098-6588 (Electronic)
IS  - 1071-412X (Linking)
VI  - 8
IP  - 3
DP  - 2001 May
TI  - Antigenic importance of the carboxy-terminal beta-strand of the porcine 
      reproductive and respiratory syndrome virus nucleocapsid protein.
PG  - 598-603
AB  - Five domains of antigenic importance were previously mapped on the nucleocapsid 
      protein (N) of the porcine reproductive and respiratory syndrome virus (PRRSV), 
      and a domain comprised of the 11 C-terminal-most amino acids (residues 112 to 
      123) was shown to be essential for binding of N-specific conformation-dependent 
      monoclonal antibodies (MAbs). In the present study, the importance of individual 
      residues within this C-terminal domain for antigenicity was investigated using 
      eight different mutant constructs of N expressed in HeLa cells. Single amino acid 
      substitutions were introduced into the C-terminal domain of the N protein, and 
      the significance of individual amino acids for MAb reactivity was determined by 
      immunoprecipitation. None of the MAbs tested recognized the mutant with a 
      leucine-to-proline substitution at residue 114 (L114P), while V112P, R113P, 
      R113D, I115P, and R116P reduced MAb binding significantly. Conversely, 
      substitution of amino acids at positions 118 (T118S) and 121 (P121A) had little 
      effect on MAb binding. Secondary-structure predictions indicate that amino acids 
      111 to 117 form a beta-strand. In view of the fact that replacement of 
      beta-strand-forming amino acids with proline elicited the greatest effect on MAb 
      binding, it appears that secondary structure in the C terminus of the N protein 
      is an important determinant of conformational epitope formation. While the 
      crystal structure of the PRRSV N protein remains to be determined, results from 
      these studies broaden our understanding of the secondary structures that make up 
      the PRRSV N protein and shed some light on how they may relate to function.
FAU - Wootton, S
AU  - Wootton S
AD  - Department of Pathobiology, Ontario Veterinary College, University of Guelph, 
      Guelph, Ontario N1G 2W1, Canada.
FAU - Koljesar, G
AU  - Koljesar G
FAU - Yang, L
AU  - Yang L
FAU - Yoon, K J
AU  - Yoon KJ
FAU - Yoo, D
AU  - Yoo D
LA  - eng
SI  - GENBANK/AF066068
SI  - GENBANK/AF092283
SI  - GENBANK/U03040
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Clin Diagn Lab Immunol
JT  - Clinical and diagnostic laboratory immunology
JID - 9421292
RN  - 0 (Antigens, Viral)
RN  - 0 (Immunodominant Epitopes)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Antigens, Viral/genetics/*immunology
MH  - Immunodominant Epitopes
MH  - Molecular Sequence Data
MH  - Nucleocapsid/genetics/*immunology
MH  - Porcine respiratory and reproductive syndrome virus/*immunology
MH  - Swine
PMC - PMC96108
EDAT- 2001/05/01 10:00
MHDA- 2001/07/20 10:01
PMCR- 2001/05/01
CRDT- 2001/05/01 10:00
PHST- 2001/05/01 10:00 [pubmed]
PHST- 2001/07/20 10:01 [medline]
PHST- 2001/05/01 10:00 [entrez]
PHST- 2001/05/01 00:00 [pmc-release]
AID - 0268 [pii]
AID - 10.1128/CDLI.8.3.598-603.2001 [doi]
PST - ppublish
SO  - Clin Diagn Lab Immunol. 2001 May;8(3):598-603. doi: 
      10.1128/CDLI.8.3.598-603.2001.