Molecular cloning and identification of a putative PKC epsilon cDNA from Limulus polyphemus brain.
The protein kinase C (PKC) family of enzymes is broadly distributed and has been implicated in a diverse array of cellular functions. Recent evidence supporting PKC involvement in the regulation of the Limulus choline cotransporter prompted us to clone PKC from a Limulus central nervous system (CNS) cDNA library. An Aplysia californica calcium independent PKC (Apl II) cDNA probe was used to screen the library and 5' RACE SMART PCR was used to obtain the full-length sequence. The resulting cDNA, which included 5' and 3' nontranslation regions, was 4675 bp. Analysis of the encoded peptide sequence using the Swiss-prot database revealed at least 58% identity to PKC epsilon. A commercial polyclonal antibody against PKC epsilon was used in Western blots to positively label a 30 kDa protein from Limulus CNS and the expressed fusion protein of the encoded sequence. These data support the presence of a newly identified PKC-like homolog in Limulus which likely represents a PKC epsilon equivalent.
Reid
Easton A
EA
Department of Anatomy and Physiology, Meharry Medical College, 1005 D.B. Todd Blvd., Nashville, TN 37208, USA.
Cao
Zheng
Z
Wang
Yilun
Y
Leite Browning
Maria L
ML
Newkirk
Robert F
RF
Chaudhuri
Guatum
G
Townsel
James G
JG
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000033:anatomy & histology / Q000201:enzymology*; D002490:Central Nervous System / Q000033:anatomy & histology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D005260:Female; D015723:Gene Library; D006737:Horseshoe Crabs / Q000033:anatomy & histology / Q000737:chemistry / Q000502:physiology*; D017403:In Situ Hybridization; D007527:Isoenzymes; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D011493:Protein Kinase C / Q000737:chemistry / Q000235:genetics*; D020539:Sequence Analysis, Protein
D018076:DNA, Complementary; D007527:Isoenzymes; D011493:Protein Kinase C