Antipeptide antibodies that can distinguish specific subunit polypeptides of glutamine synthetase from bean (Phaseolus vulgaris L.).
The amino acid sequences of the beta and gamma subunit polypeptides of glutamine synthetase from bean (Phaseolus vulgaris L.) root nodules are very similar.  However, there are small regions within the sequences that are significantly different between the two polypeptides.  The sequences between amino acids 2 and 9 and between 264 and 274 are examples.  Three peptides (gamma 2-9, gamma 264-274, and beta 264-274) corresponding to these sequences were synthesized.  Antibodies against these peptides were raised in rabbits and purified with corresponding peptide-Sepharose affinity chromatography.  Western blot analysis of polyacrylamide gel electrophoresis of bean nodule proteins demonstrated that the anti-beta 264-274 antibodies reacted specifically with the beta polypeptide and the anti-gamma 264-274 and anti-gamma 2-9 antibodies reacted specifically with the gamma polypeptide of the native and denatured glutamine synthetase.  These results showed the feasibility of using synthetic peptides in developing antibodies that are capable of distinguishing proteins with similar primary structures.
Cai
X
X
Division of Biology, Kansas State University, Manhattan 66506, USA.
Henry
R L
RL
Takemoto
L J
LJ
Guikema
J A
JA
Wong
P P
PP
D000595:Amino Acid Sequence; D000906:Antibodies; D000918:Antibody Specificity; D007887:Fabaceae / Q000201:enzymology* / Q000276:immunology; D005974:Glutamate-Ammonia Ligase / Q000032:analysis* / Q000737:chemistry; D007527:Isoenzymes; D010455:Peptides / Q000032:analysis / Q000138:chemical synthesis* / Q000737:chemistry; D018517:Plant Roots / Q000201:enzymology / Q000276:immunology; D010946:Plants, Medicinal
D000906:Antibodies; D007527:Isoenzymes; D010455:Peptides; D005974:Glutamate-Ammonia Ligase