프로테옴 분석법을 이용한 누에체액 중의 항 에이파토시스 단백질의 분리 및 동정

Abstract

The addition of silkworm hemolymph to culture medium increases the longevity of insect and mammalian cells by inhibiting apoptosis. The apoptosis-inhibiting component of silkworm hemolymph was identified as a low molecular weight lipoprotein, which is so-called '30K proteins' of unknown function. In this study, 30K proteins isolated from silkworm hemolymph by gel-filtration chromatography were studied by proteome analysis. The 30K proteins were separated into five spots, and all five spots were identified as low molecular 30kDa lipoprotein(Clone PBMHPC-19) by using MALDI-TOF. As a part of our effort to characterize the anti-apoptotic proteins in silkworm hemolymph and understand the proteome of silkworm hemolymph, silkworm hemolymph polypeptides were analyzed by 2D-gel mapping. To enhance reproducibility and resolving power of two dimensional electrophoresis, innovative method that inhibited reoxidation of –SH group was used. Compared with conventional method, in the sample treated by the new protocol a much larger number of spots is visible. The 2D-map of silkworm hemolymph was composed of over 30 Coomassie-stained protein spots. Among them, 16 spots were identified by MALDI-TOF. Silkworm hemolymph was composed of several charge isomers of differently modified isoforms of several proteins.