Soares Márcia Regina, Oliveira-Carvalho Ana Lúcia, Wermelinger Luciana Serrão, Zingali Russolina B, Ho Paulo Lee, Junqueira-de-Azevedo Inácio de L M, Diniz Marcelo Ribeiro Vasconcelos
Rede Proteômica do Rio de Janeiro and Instituto de Bioquímica Médica/CCS, Universidade Federal do Rio de Janeiro, Cidade Universitária, Rio de Janeiro 21941-590, Brazil.
Toxicon. 2005 Jul;46(1):31-8. doi: 10.1016/j.toxicon.2005.03.006.
The generation of expressed sequence tags (ESTs) from the pit-viper snake Lachesis muta venom glands allowed us to identify two cDNA isoforms which encode the precursors for bradykinin-potentiating peptides (BPPs) and a C-type natriuretic peptide (CNP). The sequence data derived from these cDNAs combined with the venom peptides identification using MALDI-TOF mass spectrometry analysis predicted that these molecules are the precursor protein isoforms that are further processed to produce five novel BPPs and a CNP. They were identified directly in crude venom using MALDI-TOF. The BPPs sequences were further confirmed by MALDI-TOF/TOF de novo sequencing, and an unusual BPP with a residue of tryptophan at the N-terminus (usually it is pyroglutamate) was identified. The putative processing steps required to form the mature BPPs and CNP seem to be similar to those proposed for the ones found in the venom of Bothrops jararaca and Glodyus blomhoffi.
从蝰蛇(Lachesis muta)毒腺中生成表达序列标签(EST),使我们能够鉴定出两种cDNA异构体,它们编码缓激肽增强肽(BPP)和C型利钠肽(CNP)的前体。这些cDNA的序列数据,结合使用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)分析进行的毒液肽鉴定,预测这些分子是前体蛋白异构体,它们会进一步加工产生五种新型BPP和一种CNP。使用MALDI-TOF在粗毒液中直接鉴定出它们。通过MALDI-TOF/TOF从头测序进一步确认了BPP的序列,并鉴定出一种在N端带有色氨酸残基(通常是焦谷氨酸)的异常BPP。形成成熟BPP和CNP所需的假定加工步骤似乎与拟眼镜蛇(Bothrops jararaca)和日本蝮蛇(Glodyus blomhoffi)毒液中发现的那些相似。