Antiendotoxin agents share molecular homology within their lipopolysaccharide binding domains.
BACKGROUND
The purpose of this study was to determine whether antiendotoxin agents exhibit molecular homology within their lipopolysaccharide (LPS) binding domains, suggesting a common mechanism of action. We hypothesized that the presence of positively charged basic amino acids or a paucity of negatively charged acidic amino acids, or both, would be a critical characteristic of that portion of the molecule that binds to the highly negatively charged deep core/lipid A (DCLA) region of LPS.


METHODS
We analyzed the amino acid sequences of the variable light (VL) and heavy (VH) chain complementarity-determining regions (CDRs) of anti-DCLA monoclonal antibodies (mAbs) 1B6, 5A5, and 7C5 and compared them with (1) the CDRs of three irrelevant control mAbs and (2) the LPS binding region of bactericidal permeability-increasing protein (BPI). We purified and amplified the specific nucleotide sequences of the variable regions using reverse transcriptase polymerase chain reaction. DNA was sequenced by dideoxy termination, and protein sequences were deduced and analyzed. The percentages of acidic, basic, polar, and hydrophobic amino acids within VH and VL chain CDRs were determined.


RESULTS
We identified a paucity of negatively charged acidic amino acids exclusively within VL chain CDRs of anti-DCLA mAbs (P < 0.005). Although increased, the number of positively charged basic residues was not statistically significantly different; neither was the number of polar or hydrophobic amino acids. Conclusions. Our data suggest that the near absence of negatively charged acidic residues is critical for LPS binding. This characteristic appears to reside exclusively in the VL chain CDRs of anti-DCLA mAbs.
Kellogg
T A
TA
Department of Surgery, University of Minnesota, Minneapolis, Minnesota, 55455, USA.
Weiss
C A
CA
Johnston
J W
JW
Wasiluk
K R
KR
Dunn
D L
DL
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000494:pharmacology; D004731:Endotoxins / Q000037:antagonists & inhibitors* / Q000276:immunology; D005260:Female; D007135:Immunoglobulin Variable Region / Q000737:chemistry; D008070:Lipopolysaccharides / Q000737:chemistry* / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D051381:Rats; D017386:Sequence Homology, Amino Acid
D000911:Antibodies, Monoclonal; D004731:Endotoxins; D007135:Immunoglobulin Variable Region; D008070:Lipopolysaccharides