Island specific expression of a novel [Lys(49)]phospholipase A(2) (BPIII) in Protobothrops flavoviridis venom in Amami-Oshima, Japan.
In search of the transcripts expressed in Protobothrops flavoviridis venom gland, 466 expressed sequence tags (ESTs) were generated from the venom gland cDNA library of P. flavoviridis in Amami-Oshima, Japan. The sequencing of randomly selected cDNA clones followed by identification in similarity search against existing databases led to the finding of a novel lysine-49-phospholipase A(2) ([Lys(49)]PLA(2)) clone. It coded for one amino acid-substituted BPII homologue or two amino acids-substituted BPI homologue in which BPII and BPI are [Lys(49)]PLA(2)s contained in Amami-Oshima and Tokunoshima P. flavoviridis venoms. This isozyme, named BPIII, was isolated from Amami-Oshima P. flavoviridis venom. BPIII gave a specific [M+2H](2+) peak of m/z 736.3 on mass spectrometry (MS) analysis after S-carboxamidomethylation and trypsin digestion when compared with BPII. It became evident from MS analysis after S-carboxamidomethylation and trypsin digestion of the mixed protein peaks ranging from BPI to BPII obtained by fractionation on a carboxymethyl cellulose column of Amami-Oshima and Tokunoshima P. flavoviridis venoms that BPIII protein is contained in Amami-Oshima P. flavoviridis venom but not in Tokunoshima P. flavoviridis venom. It is for the first time that a protein present in Amami-Oshima P. flavoviridis venom is not found in Tokunoshima P. flavoviridis venom.
Murakami
Tatsuo
T
Department of Bioscience and Biotechnology, Sojo University, 4-22-1 Ikeda, Kumamoto 860-0082, Japan.
Kariu
Toru
T
Takazaki
Shinya
S
Hattori
Shosaku
S
Chijiwa
Takahito
T
Ohno
Motonori
M
Oda-Ueda
Naoko
N
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D000925:Anticoagulants / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D001483:Base Sequence; D005591:Chemical Fractionation; D003435:Crotalid Venoms / Q000737:chemistry / Q000201:enzymology*; D020224:Expressed Sequence Tags; D015723:Gene Library; D005843:Geography; D006801:Humans; D007564:Japan; D019169:Jurkat Cells; D008239:Lysine / Q000737:chemistry; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D054467:Phospholipases A2 / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D016415:Sequence Alignment; D017819:Viperidae / Q000378:metabolism*
D000925:Anticoagulants; D003435:Crotalid Venoms; D054467:Phospholipases A2; D008239:Lysine