Specific chaperone-like activity of inhibitor of caspase-activated DNase for caspase-activated DNase.
Caspase-activated DNase (CAD) is the enzyme that causes DNA fragmentation during apoptosis. CAD forms aggregates when it is synthesized in the absence of an inhibitor of CAD (ICAD). Here, using renaturation systems of chemically denatured CAD, we report that ICAD-L, a long form of ICAD, has a chaperone-like activity specific for CAD. Murine CAD carries 14 cysteines, most of which were found to be in reduced form. Reducing agents enhanced the production of the functional CAD in an in vitro translation system. The denatured CAD could be efficiently renatured under highly reducing conditions only in the presence of ICAD-L. This process was ATP-independent. In contrast, reticulocyte lysates stimulated ICAD-L- and ATP-dependent renaturation of denatured CAD without requiring a high concentration of reducing agents. These results indicate that ICAD-L works not only as a specific inhibitor but also as a specific chaperone for CAD.
Sakahira
H
H
Department of Genetics, Osaka University Medical School, Japan Science and Technology Corporation, 2-2 Yamada-oka, Suita, Osaka 565-0871, Japan.
Iwamatsu
A
A
Nagata
S
S
D000478:Alkylation; D000595:Amino Acid Sequence; D000818:Animals; D051017:Apoptosis Regulatory Proteins; D002474:Cell-Free System; D003851:Deoxyribonucleases / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D013058:Mass Spectrometry; D051379:Mice; D018832:Molecular Chaperones / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D014176:Protein Biosynthesis; D011489:Protein Denaturation; D017510:Protein Folding; D011506:Proteins / Q000378:metabolism*; D019163:Reducing Agents
D051017:Apoptosis Regulatory Proteins; D018832:Molecular Chaperones; D010446:Peptide Fragments; D011506:Proteins; D019163:Reducing Agents; C109664:caspase-activated DNase inhibitor; D003851:Deoxyribonucleases; C109663:caspase-activated deoxyribonuclease