PMID- 1939118
OWN - NLM
STAT- MEDLINE
DCOM- 19911213
LR  - 20210210
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 266
IP  - 31
DP  - 1991 Nov 5
TI  - Physical characterization and crystallization of the carbohydrate-recognition 
      domain of a mannose-binding protein from rat.
PG  - 20678-86
AB  - A portion of rat mannose-binding protein A (MBP-A), a Ca(2+)-dependent animal 
      lectin, has been overproduced in a bacterial expression system, biochemically 
      characterized, and crystallized. A fragment corresponding to the COOH-terminal 
      115 residues of native MBP-A, produced by subtilisin digestion of the bacterially 
      expressed protein, contains the carbohydrate-recognition domain (CRD). Gel 
      filtration, chemical cross-linking, and crystallographic self-rotation function 
      analyses indicate that the subtilisin fragment is a dimer, although the complete 
      bacterially expressed fragment, containing the neck and CRD of MBP-A, is a 
      trimer. Crystals of the minimal CRD, obtained only as a complex with a 
      Man6GlcNAc2Asn glycopeptide, diffract to Bragg spacings of at least 1.7 A. 
      Several trivalent lanthanide ions (Ln3+) can substitute for Ca2+, as assessed by 
      their ability to support carbohydrate binding and to protect the CRD from 
      proteolysis in a manner similar to that observed for Ca2+. These assays indicate 
      that Ln2+ binds about 30 times more tightly than Ca2+ to the CRD, and that two 
      Ca2+ or Ln3+ bind to each monomer, a result confirmed by determination of the 
      Ho3+ positions in a Ho(3+)-containing crystal of the CRD. Crystals grown in the 
      presence of Ln3+ belong to different space groups from those obtained with Ca2+ 
      and are therefore not useable for traditional crystallographic phase 
      determination methods, but are well-suited for high resolution structure 
      determination by multiwavelength anomalous dispersion phasing.
FAU - Weis, W I
AU  - Weis WI
AD  - Department of Biochemistry and Molecular Biophysics, Columbia University, New 
      York, New York 10032.
FAU - Crichlow, G V
AU  - Crichlow GV
FAU - Murthy, H M
AU  - Murthy HM
FAU - Hendrickson, W A
AU  - Hendrickson WA
FAU - Drickamer, K
AU  - Drickamer K
LA  - eng
GR  - GM34102/GM/NIGMS NIH HHS/United States
GR  - GM42628/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Carrier Proteins)
RN  - 0 (Cations)
RN  - 0 (Lectins)
RN  - 0 (Mannose-Binding Lectins)
RN  - 0 (Peptide Fragments)
RN  - 0 (Recombinant Proteins)
RN  - 6I3K30563S (Lanthanum)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Animals
MH  - Binding Sites
MH  - Calcium/chemistry
MH  - Carbohydrate Sequence
MH  - Carrier Proteins/*chemistry
MH  - Cations
MH  - Chromatography, Gel
MH  - Crystallography
MH  - DNA Mutational Analysis
MH  - Lanthanum/chemistry
MH  - Lectins/*chemistry
MH  - Mannose-Binding Lectins
MH  - Molecular Sequence Data
MH  - Molecular Weight
MH  - Peptide Fragments/chemistry
MH  - Peptide Mapping
MH  - Rats
MH  - Recombinant Proteins/metabolism
MH  - Structure-Activity Relationship
EDAT- 1991/11/05 00:00
MHDA- 1991/11/05 00:01
CRDT- 1991/11/05 00:00
PHST- 1991/11/05 00:00 [pubmed]
PHST- 1991/11/05 00:01 [medline]
PHST- 1991/11/05 00:00 [entrez]
AID - S0021-9258(18)54762-1 [pii]
PST - ppublish
SO  - J Biol Chem. 1991 Nov 5;266(31):20678-86.