PMID- 11350170
OWN - NLM
STAT- MEDLINE
DCOM- 20010607
LR  - 20131121
IS  - 0022-2836 (Print)
IS  - 0022-2836 (Linking)
VI  - 308
IP  - 4
DP  - 2001 May 11
TI  - Structure and binding determinants of the recombinant kringle-2 domain of human 
      plasminogen to an internal peptide from a group A Streptococcal surface protein.
PG  - 705-19
AB  - The X-ray crystal structure of a complex of a modified recombinant kringle-2 
      domain of human plasminogen, K2Pg[C4G/E56D/L72Y] (mK2Pg), containing an 
      upregulated lysine-binding site, bound to a functional 30 residue internal 
      peptide (VEK-30) from an M-type protein of a group A Streptococcus surface 
      protein, has been determined by molecular replacement methods using K4Pg as a 
      model, and refined at 2.7 A resolution to a R-factor of 19.5 %. The X-ray crystal 
      structure shows that VEK-30 exists as a nearly end-to-end alpha-helix in the 
      complex with mK2Pg. The final structure also revealed that Arg17 and His18 of 
      VEK-30 served as cationic loci for Asp54 and Asp56 of the consensus 
      lysine-binding site of mK2Pg, while Glu20 of VEK-30 coordinates with Arg69 of the 
      cationic binding site of mK2Pg. The hydrophobic ligand-binding pocket in mK2Pg, 
      consisting primarily of Trp60 and Trp70, situated between the positive and 
      negative centers of the lysine-binding site, is utilized in a novel manner in 
      stabilizing the interaction with VEK-30 by forming a cation-pi-electron-mediated 
      association with the positive side-chain of Arg17 of this peptide. Additional 
      lysine-binding sites, as well as exosite electrostatic and hydrogen bonding 
      interactions involving Glu9 and Lys14 of VEK-30, were observed in the structural 
      model. The importance of these interactions were tested in solution by 
      investigating the binding constants of synthetic variants of VEK-30 to mK2Pg, and 
      it was found that, Lys14, Arg17, His18, and Glu20 of VEK-30 were the most 
      critical amino acid binding determinants. With regard to the solution studies, 
      circular dichroism analysis of the titration of VEK-30 with mK2Pg demonstrated 
      that the peptidic alpha-helical structure increased substantially when bound to 
      the kringle module, in agreement with the X-ray results. This investigation is 
      the first to delineate structurally the mode of interaction of the lysine-binding 
      site of a kringle with an internal pseudo-lysine residue of a peptide or protein 
      that functionally interacts with a kringle module, and serves as a paradigm for 
      this important class of interactions.
CI  - Copyright 2001 Academic Press.
FAU - Rios-Steiner, J L
AU  - Rios-Steiner JL
AD  - Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, 
      IN 46556, USA.
FAU - Schenone, M
AU  - Schenone M
FAU - Mochalkin, I
AU  - Mochalkin I
FAU - Tulinsky, A
AU  - Tulinsky A
FAU - Castellino, F J
AU  - Castellino FJ
LA  - eng
GR  - HL13423/HL/NHLBI NIH HHS/United States
GR  - HL25942/HL/NHLBI NIH HHS/United States
GR  - HL43229/HL/NHLBI NIH HHS/United States
GR  - RR07707/RR/NCRR NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - Netherlands
TA  - J Mol Biol
JT  - Journal of molecular biology
JID - 2985088R
RN  - 0 (Antigens, Bacterial)
RN  - 0 (Ligands)
RN  - 0 (Peptide Fragments)
RN  - 0 (Recombinant Proteins)
RN  - 9001-91-6 (Plasminogen)
RN  - K3Z4F929H6 (Lysine)
SB  - IM
EIN - J Mol Biol 2001 Sep 14;312(2):421
MH  - Amino Acid Sequence
MH  - Amino Acid Substitution/genetics
MH  - Antigens, Bacterial/*chemistry/genetics/*metabolism
MH  - Binding Sites
MH  - Calorimetry
MH  - Circular Dichroism
MH  - Crystallography, X-Ray
MH  - Humans
MH  - Hydrogen Bonding
MH  - *Kringles
MH  - Ligands
MH  - Lysine/metabolism
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Peptide Fragments/chemistry/genetics/*metabolism
MH  - Plasminogen/*chemistry/*metabolism
MH  - Protein Binding
MH  - Protein Structure, Secondary
MH  - Recombinant Proteins/chemistry/metabolism
MH  - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
MH  - Static Electricity
MH  - Streptococcus/*chemistry/genetics
MH  - Thermodynamics
EDAT- 2001/05/15 10:00
MHDA- 2001/06/08 10:01
CRDT- 2001/05/15 10:00
PHST- 2001/05/15 10:00 [pubmed]
PHST- 2001/06/08 10:01 [medline]
PHST- 2001/05/15 10:00 [entrez]
AID - S0022-2836(01)94646-7 [pii]
AID - 10.1006/jmbi.2001.4646 [doi]
PST - ppublish
SO  - J Mol Biol. 2001 May 11;308(4):705-19. doi: 10.1006/jmbi.2001.4646.