Primary structure, isoforms, and molecular modeling of a chitin-binding mistletoe lectin.
From mistletoe Viscum album L. extracts three chitin-binding lectin isoforms, cbML1, cbML2, and cbML3, were isolated and their primary structure determined. All three cbML isoforms are composed of two protein chains of 48 or 49 amino acid residues, linked by an intermolecular disulfide bond. The sequence of each single cbML chain is characterized by a relatively high number of cysteine and glycine residues, 9 and 6, respectively, and contains four intramolecular disulfide bridges. On the basis of the combined interpretation of sequencing and MALDI MS data, the following results for the three cbML isoforms were obtained: the first one consists of two identical truncated polypeptide chains (1--48), the second is a heterodimer, containing one truncated (1--48) and one full-length chain (1--49), and the third is composed of two full length chains (1--49). The cbML sequence shows 55% identity to hevein, a single-chain chitin-binding protein of 43 amino acids, one of the most predominant proteins in natural rubber latex. On the basis of the NMR data on hevein from Hevea brasiliensis the three-dimensional structure of cbML3 was modelled. The 26 sequence changes between cbML3 and hevein were accommodated with only little perturbation in the main chain folding. A comparison of the primary structures of cbML3 and hevein is shown and the effects of the sequence changes are discussed. Differences have been identified in the loop region of the molecule and the potential interface region of cbML3 supporting the dimer formation. The high-affinity chitin-binding site seems to be highly conserved.
Stoeva
S
S
Department of Physical Biochemistry, Institute of Physiological Chemistry, University of Tuebingen, Hoppe-Seyler-Strasse 4, D-72076 Tuebingen, Germany. stanka.stoeva@uni-tuebingen.de
Franz
M
M
Wacker
R
R
Krauspenhaar
R
R
Guthöhrlein
E
E
Mikhailov
A
A
Betzel
C
C
Voelter
W
W
D000595:Amino Acid Sequence; D002686:Chitin / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D019281:Dimerization; D014784:Mistletoe / Q000737:chemistry* / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D028321:Plant Preparations; D010940:Plant Proteins; D010946:Plants, Medicinal; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D020836:Protein Structure, Quaternary; D054790:Ribosome Inactivating Proteins, Type 2; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014118:Toxins, Biological / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D028321:Plant Preparations; D010940:Plant Proteins; D020033:Protein Isoforms; D054790:Ribosome Inactivating Proteins, Type 2; D014118:Toxins, Biological; C009674:ribosome inactivating protein, Viscum; D002686:Chitin