Enzymatic properties of sialic acid binding lectin from Rana catesbeiana modified with a water-soluble carbodiimide in the presence of various nucleophiles.
The anti-tumor activity of sialic acid binding lectin from Rana catesbeiana (cSBL) was increased by chemical modification with a water-soluble carbodiimide (EDC) in the presence of nucleophiles such as ethylenediamine and glycine methylester. Investigations on ribonuclease (RNase) activities of the modified cSBLs were conducted to elucidate the fundamental mechanisms underlying enhancement of the anti-tumor activity conferred by these modifications. The following three characteristics were observed with modification. (i) RNase activity of the modified cSBL was enhanced towards double stranded RNA and RNA-oligo dA hybrids. The activity increase was observed even under physiologic ionic strength conditions; (ii) RNase activity of the modified cSBL towards single stranded RNA and poly U decreased, while the activity towards poly C was unaffected; (iii) the base preference of the B2 base recognition site of modified cSBL decreased for guanine. On the contrary, the preference for cytosine and adenine increased. This result may explain why the RNase activity towards poly C was not affected by EDC-modification as mentioned above.
Iwama
M
M
Department of Microbiology Hoshi College of Pharmacy, Tokyo, Japan. iwama@nagaoka-ct.ac.jp
Ogawa
Y
Y
Ohgi
K
K
Tsuji
T
T
Irie
M
M
D029845:Amphibian Proteins; D000818:Animals; D015226:Dinucleoside Phosphates / Q000378:metabolism; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology*; D006868:Hydrolysis; D037102:Lectins / Q000378:metabolism*; D011072:Poly U / Q000378:metabolism; D011684:Purine Nucleosides / Q000378:metabolism* / Q000494:pharmacology; D011741:Pyrimidine Nucleosides / Q000378:metabolism* / Q000494:pharmacology; D012330:RNA, Double-Stranded / Q000378:metabolism; D011892:Rana catesbeiana; D012260:Ribonucleases / Q000378:metabolism*; D012995:Solubility; D014529:Uridine / Q000378:metabolism
D029845:Amphibian Proteins; D015226:Dinucleoside Phosphates; D037102:Lectins; D011684:Purine Nucleosides; D011741:Pyrimidine Nucleosides; D012330:RNA, Double-Stranded; D011072:Poly U; C041829:1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide methiodide; D012260:Ribonucleases; C448662:leczyme protein, Rana; D005022:Ethyldimethylaminopropyl Carbodiimide; D014529:Uridine