Cloning and expression of sialidase L, a NeuAc alpha 2->3Gal-specific sialidase from the leech, Macrobdella decora

被引:24
作者
Chou, MY [1 ]
Li, SC [1 ]
Li, YT [1 ]
机构
[1] TULANE UNIV,SCH MED,DEPT BIOCHEM,NEW ORLEANS,LA 70112
关键词
D O I
10.1074/jbc.271.32.19219
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Sialidase L is a NeuAc alpha 2-->3Gal linkage-specific sialidase that releases 2,7-anhydro-NeuAc instead of NeuAc from sialoglycoconjugates (Chou, M.-Y., Li, S.-C., Kiso, M,, Hasegawa, A., and Li, Y.-T, (1994) J. Biol, Chem. 269, 18821-18826), A 2.5-kilobase cDNA of sialidase L was cloned by a combination of methods based on polymerase chain reactions, The composite cDNA sequence reveals an open reading frame coding for 762 amino acids, including a putative 28-residue signal peptide at the N terminus that is similar to the signal sequence of the Clostridium septicum sialidase, The result suggests that sialidase L is a secretory enzyme, The coding sequence excluding the putative signal peptide of sialidase L was overexpressed in Escherichia coli. The purified recombinant enzyme was characterized to be as active as the enzyme isolated from the leech, It also possessed the strict NeuAc alpha 2-->3Gal linkage specificity and released the unique cleavage product, 2,7-anhydro-NeuAc from sialoglycoconjugates. The deduced amino acid sequence of sialidase L exhibits little similarity with other reported sialidases, However, sialidase L contains a conserved ''FRIP region'' and four repeating ''Asp box'' motifs that align well with the corresponding positions of bacterial sialidases. The predicted beta-strand structures near the conserved motifs of sialidase L are similar to those of Salmonella typhimurium sialidase. Several conserved single amino acid residues of bacterial sialidases, including those known to be involved in the active site of Salmonella enzyme, are conserved in the deduced amino acid sequence of sialidase L, This observation suggests that part of the catalytic mechanism of sialidase L may be similar to the ordinary sialidase.
引用
收藏
页码:19219 / 19224
页数:6
相关论文
共 31 条
[1]   COMPLETE SEQUENCE OF THE BACTEROIDES-FRAGILIS YCH46 NEURAMINIDASE-ENCODING GENE [J].
AKIMOTO, S ;
ONO, T ;
TSUTSUI, H ;
KINOUCHI, T ;
KATAOKA, K ;
OHNISHI, Y .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1994, 203 (02) :914-921
[2]  
CAMARA M, 1994, INFECT IMMUN, V62, P3688
[3]  
CHOU MY, 1994, J BIOL CHEM, V269, P18821
[4]   CRYSTAL-STRUCTURE OF A BACTERIAL SIALIDASE (FROM SALMONELLA-TYPHIMURIUM LT2) SHOWS THE SAME FOLD AS AN INFLUENZA-VIRUS NEURAMINIDASE [J].
CRENNELL, SJ ;
GARMAN, EF ;
LAVER, WG ;
VIMR, ER ;
TAYLOR, GL .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (21) :9852-9856
[5]   CLONING AND EXPRESSION OF A SOLUBLE SIALIDASE FROM CHINESE-HAMSTER OVARY CELLS - SEQUENCE ALIGNMENT SIMILARITIES TO BACTERIAL SIALIDASES [J].
FERRARI, J ;
HARRIS, R ;
WARNER, TG .
GLYCOBIOLOGY, 1994, 4 (03) :367-373
[6]   RAPID PRODUCTION OF FULL-LENGTH CDNAS FROM RARE TRANSCRIPTS - AMPLIFICATION USING A SINGLE GENE-SPECIFIC OLIGONUCLEOTIDE PRIMER [J].
FROHMAN, MA ;
DUSH, MK ;
MARTIN, GR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (23) :8998-9002
[7]   ANALYSIS OF ACCURACY AND IMPLICATIONS OF SIMPLE METHODS FOR PREDICTING SECONDARY STRUCTURE OF GLOBULAR PROTEINS [J].
GARNIER, J ;
OSGUTHORPE, DJ ;
ROBSON, B .
JOURNAL OF MOLECULAR BIOLOGY, 1978, 120 (01) :97-120
[8]   ISOLATION OF FULL-LENGTH PUTATIVE RAT LYSOPHOSPHOLIPASE CDNA USING IMPROVED METHODS FOR MESSENGER-RNA ISOLATION AND CDNA CLONING [J].
HAN, JH ;
STRATOWA, C ;
RUTTER, WJ .
BIOCHEMISTRY, 1987, 26 (06) :1617-1625
[9]  
HARRIS G, 1954, CHEM IND-LONDON, P249
[10]   CLONING, SEQUENCING AND EXPRESSION OF THE SIALIDASE GENE FROM ACTINOMYCES-VISCOSUS DSM 43798 [J].
HENNINGSEN, M ;
ROGGENTIN, P ;
SCHAUER, R .
BIOLOGICAL CHEMISTRY HOPPE-SEYLER, 1991, 372 (12) :1065-1072