Crocus sativus lectin recognizes Man3GlcNAc in the N-glycan core structure.
Crocus sativus lectin (CSL) is one of the truly mannose-specific plant lectins that has a unique binding specificity that sets it apart from others. We studied sugar-binding specificity of CSL in detail by a solution phase method (fluorescence polarization) and three solid phase methods (flow injection, surface plasmon resonance, and microtiter plate), using a number of different glycopeptides and oligosaccharides. CSL binds the branched mannotriose structure in the N-glycan core. Substitution of the terminal Man in the Manalpha(1-3)Man branch with GlcNAc drastically decreases binding affinity much more than masking of the terminal Man in the Manalpha(1-6)Man branch. Most interestingly, the beta-Man-linked GlcNAc in N-glycan core structure contributes greatly to the binding. The effect of this GlcNAc is so strong that it can substantially offset the negative effect of substitution on the nonreducing terminal Man residues. On the other hand, the GlcNAc that is usually attached to Asn in N-glycans and the l-Fuc linked at the 6-position of the GlcNAc are irrelevant to the binding. A bisecting GlcNAc neither contributes to nor interferes with the binding. This unique binding specificity of CSL offers many possibilities of its use in analytical and preparative applications.
Oda
Y
Y
Faculty of Pharmaceutical Sciences, Kinki University, Kowakae 3-4-1, Higashi-Osaka 577-8502 Japan.
Nakayama
K
K
Abdul-Rahman
B
B
Kinoshita
M
M
Hashimoto
O
O
Kawasaki
N
N
Hayakawa
T
T
Kakehi
K
K
Tomiya
N
N
Lee
Y C
YC
D000818:Animals; D001665:Binding Sites; D001704:Biopolymers; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D003370:Coturnix; D005454:Fluorescence Polarization; D037102:Lectins / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*; D011485:Protein Binding; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D001704:Biopolymers; D037102:Lectins; D009844:Oligosaccharides; D011134:Polysaccharides