[Study of the structure-activity features of lactate dehydrogenase isoenzymes using antipeptide antibodies to the M4-isoform of swine lactate dehydrogenase].
The structure-function peculiarities of human, porcine, rabbit, and rat lactate dehydrogenase (LDH) isoenzymes have been studied using antipeptide antibodies (AB) against the M4-isoform of porcine LDH. Antipeptide AB were raised against the hypervariable (40% homology) N-terminal fragment (residues 1-32), and the highly conservative fragment 180-214 containing histidine in the enzyme active center. Whereas antipeptide AB against the fragment of the active center of porcine LDH M4-isoform selectively inhibited the catalytic activities of LDH isoenzymes from various sources, antipeptide AB directed against the N-end were without effect. The ability of antipeptide AB to specifically interact with various isoforms of LDH suggests that sequences 1-32 and 180-214 are immunochemically identical only in the case of human and porcine M4 isoenzymes; the relatedness of the amino acid sequence to the common antigenic determinant required the absence in the given sequence of essential amino acid substituents. Chemical modification of porcine M4-isoform by diethylpyrocarbonate and the use of specific AB revealed that histidine-195 located in the active center of LDH is not directly involved in the binding to AB.
Grebenshchikova
O G
OG
Alekseeva
A E
AE
Potapkina
T A
TA
Prozorovskiĭ
V N
VN
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D001665:Binding Sites; D004587:Electrophoresis, Agar Gel; D006801:Humans; D007150:Immunohistochemistry; D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology*; D011817:Rabbits; D051381:Rats; D013045:Species Specificity; D013329:Structure-Activity Relationship; D013552:Swine
D000906:Antibodies; D007527:Isoenzymes; D010455:Peptides; D007770:L-Lactate Dehydrogenase
Issledovanie strukturno-funktsional'nykh osobennosteĭ izofermentov laktatdegidrogenazy s pomoshchĭu antipeptidnykh antitel k M4-izoforme laktatdegidrogenazy svin'i.