Identification of a nucleic acid helix-destabilizing protein from rat liver as lactate dehydrogenase-5.
A rat liver DNA helix-destabilizing protein (HDP) that has previously been proposed to play a role in transcription has been identified as M chain lactate dehydrogenase (LDH-5; L-lactate:NAD+ oxidoreductase, EC 1.1.1.27). Tryptic peptides accounting for 157 amino acids in the rat liver HDP have been characterized and then matched to the published sequence for the M chain of porcine LDH. Based on amino acid compositions and direct solid-phase protein sequencing, at least 148 of the 157 residues that were compared are identical in both proteins. In addition, both porcine LDH and the rat liver HDP have blocked amino termini and similar amino acid compositions and molecular weights. Rat liver HDP and LDH-5 that were purified to molecular homogeneity had similar specific activities in both single-stranded DNA (ss DNA) binding and LDH assays. HPLC tryptic peptide maps were also identical for both the rat liver HDP and LDH proteins. Since preincubation of HDP in NADH prevents its binding to ss DNA, both NADH and ss DNA may be binding at the same site. Further support for this latter idea derives from chemical-modification studies which demonstrate that tyrosine-238, which is located near the coenzyme binding site of LDH, seems to be essential for the ability of HDP to bind ss DNA. These results indicate caution in ascribing in vivo roles solely on the basis of binding to ss DNA. Alternatively, they suggest that a single protein may play more than one biological role.
Williams
K R
KR
Reddigari
S
S
Patel
G L
GL
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004265:DNA Helicases / Q000302:isolation & purification / Q000378:metabolism*; D004268:DNA-Binding Proteins; D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase / Q000302:isolation & purification / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D010446:Peptide Fragments / Q000032:analysis; D051381:Rats; D012261:Ribonucleoproteins / Q000378:metabolism*; D013552:Swine; D014357:Trypsin; D014764:Viral Proteins
D004268:DNA-Binding Proteins; D007527:Isoenzymes; D010446:Peptide Fragments; D012261:Ribonucleoproteins; D014764:Viral Proteins; C060035:gp32 protein, Enterobacteria phage T4; C027729:helix-destabilizing proteins; D007770:L-Lactate Dehydrogenase; D014357:Trypsin; D004265:DNA Helicases