 |
PDBsum entry 2hwz
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Immune system
|
PDB id
|
|
|
|
2hwz
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
References listed in PDB file
|
|
|
Key reference
|
|
|
Title
|
|
Identification of a single tryptophan residue as critical for binding activity in a humanized monoclonal antibody against respiratory syncytial virus.
|
|
|
Authors
|
|
Z.Wei,
J.Feng,
H.Y.Lin,
S.Mullapudi,
E.Bishop,
G.I.Tous,
J.Casas-Finet,
F.Hakki,
R.Strouse,
M.A.Schenerman.
|
|
|
Ref.
|
|
Anal Chem, 2007,
79,
2797-2805.
|
|
|
PubMed id
|
|
|
|
|
|
Abstract
|
|
|
We have identified a single tryptophan (Trp) residue responsible for loss of
binding and biological activity upon ultraviolet (UV) light irradiation in
MEDI-493, a humanized monoclonal antibody (MAb) against respiratory syncytial
virus (RSV). This finding provides a better understanding of structure-function
relationship in a 150-kDa protein. Irradiation of MEDI-493 with UV light
resulted in spectral changes typical of Trp photoproducts and in a progressive
loss of MEDI-493 binding and biological activity as measured by ELISA, Biacore,
and cell-based assays. Mass spectrometric characterization of the proteolytic
peptides generated from the UV irradiated MEDI-493 confirmed that most
methionine (Met) and a few Trp residues were oxidized to various extents upon
exposure to UV light. Among Trp residues, only Trp-105, containing the most
solvent-exposed indole moiety in MEDI-493 and residing in a
complementary-determining region (CDR) of the heavy chain, was significantly
oxidized. When bound to a synthetic antigenic peptide, MEDI-493 showed
significant resistance toward binding activity loss during UV irradiation. A
second MAb (MEDI-524) with Trp-105 replaced by phenylalanine (Phe) showed a
similar pattern of Met oxidation, but no loss of binding and biological activity
following irradiation. Treatment of both MAbs with Met- and Trp-specific
oxidizing reagents showed that oxidation of Trp-105 correlated with the activity
loss, whereas Met oxidation did not affect the activity. These results
demonstrate that Trp-105 in MEDI-493 is responsible for the UV light-induced
effects.
|
|
|
Secondary reference #1
|
|
|
Title
|
|
Identification of a single tryptophan residue as critical for binding activity in a humanized monoclonal antibody against respiratory syncytial virus.
|
|
|
Authors
|
|
Z.Wei,
J.Feng,
H.Y.Lin,
S.Mullapudi,
E.Bishop,
G.I.Tous,
J.Casas-Finet,
F.Hakki,
R.Strouse,
M.A.Schenerman.
|
|
|
Ref.
|
|
Anal Chem, 2007,
79,
2797-2805.
|
|
|
PubMed id
|
|
|
|
|
Note In the PDB file this reference is
annotated as "TO BE PUBLISHED".
The citation details given above were identified by an automated
search of PubMed on title and author
names, giving a
perfect match.
|
|
|
|
|
|
|
|
|
|
|
