Molecular cloning and characterization of rat and human calpain-5.
Until today, 14 isoforms of mammalian calpains have been identified, including calpain-5. The C. elegans calpain-5 homologue tra-3 is reported to be essential for necrotic neuronal cell death. In this study, we cloned and characterized rat calpain-5, which is highly homologous to human and mouse sequences. The nucleotide sequence is 87% and 93% identical with human and mouse calpain-5, respectively. The protein sequence is well conserved, showing 96% identity in mouse and 92% in human. RT-PCR analysis revealed strong expression of calpain-5 in rat lungs, kidneys, and brain while week expression in heart, whereas in rat brain regions it is ubiquitously expressed. The mRNA expression in different human tissues showed equal expression. However, in human brain regions calpain-5 was strongly expressed in hypothalamus, thalamus, cerebellum, and frontal lobe. Western blot analysis on human neuroblastoma SH-SY5Y cells demonstrated calcium-dependent processing of calpain-5, despite the absence of calmodulin-like domain.
Department of Psychiatry, McKnight Brain Institute of the University of Florida, 100 S Newell Drive, P.O. Box 100256, Gainesville, FL 32610-0256, USA. anu@mbi.ufl.edu <anu@mbi.ufl.edu>
Anuradhaa
A
Waghray
Deng-shun
DS
Wang
Deborah
D
McKinsey
Ronald L
RL
Hayes
Kevin K W
KK
Wang
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002154:Calpain / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014018:Tissue Distribution
D020033:Protein Isoforms; D002154:Calpain; C494426:Capn5 protein, human; C403235:Capn5 protein, mouse; C494425:calpain 5, rat