A   Tachylectin-2 Structure and Function
1.) Introductory text on invertebrate lectins and cognate carbohydrates, bringing up the main topics of the underlying EMBO paper (tachylectin-2) 
2.) Description of the tachylectin-2 structure as a unique 5-bladed propeller structure.
3.) Presentation of images and models of the tachylectin-2 molecule in complex with GlucNAc.
4.) Presentation of tachylectin-2 still images generated with rasmol.
5.) Presentation of structural descriptions and definitions of tachylectin-2 generated from    the  SCOP and CATH databases; structural neighbour analysis, and ClustalW analysis.
6.) Description and definitions of other propeller proteins with different symmetries.
7.) Discussion of structure-function relationship of distinct propeller elements in ligand recognition.
8.) Presentation of images and models of the tachylectin-5A molecule in complex with carbohydrate ligand (PNAS paper).

B Technical Section on Tachylectin Crystallography
1.) Materials and Methods section from the EMBO paper; explanation of the crystallization strategy and the steps of data analysis taken.
2.) Technical comments from the crystallography segments of deposited tachylectin files.
3.) Explanation of some advanced features used in the tachylectin-2 structural analysis, such as the Wilson plot (correction diagram for temperature-dependent atomic vibrations).

Click here to go to the page describing the structure of tachylectin-5A.

A2 Tachylectin-2 Structure: 
a unique 5-bladed Propeller 
The overall shape of tachylectin-2 is that of a torus consisting of five almost identical structural elements, built mostly from 4 antiparallel b-sheets, that are organized around a central tunnel. The height of the torus is about 25 Angstrom, the apparent diameter is 45-48 Angstrom, and the width of the central tunnel is less than 2 Angstrom, if the water molecules are considered. 

Each of the 5 b-sheets adopts a W-like topology, thus resembling in shape the individual blade of a propeller formed by the sum of the b sheets.The resemblance with a propeller has lend the name to the defining structural feature of a large and highly diversified group of proteins that show blade symmetries ranging between 4 and 8.
(Click here to go to the page describing those other propeller symmetries and discussing some functional roles of b-propeller domains).
In tachylectin-2, the blades are numbered from I to V. When superimposed, the five b-sheets reveal their structural equivalence with a mean r.m.s. distance of 0.46 Angstrom. Eachb sheet is numbered such that it begins with strand 1 abutting the central tunnel of the molecule, whereas strand 4 is the outermost strand that forms part of the outer surface. The spacer length between the antiparallel strands is variable within a given sheet. Between strands 1 and 2, five amino acids form the spacer; strand 2 and 3 are connected very tightly by a b-turn consisting of two amino acids. Strands 3 and 4 are connected by a large loop subdivided into three structural elements: 9 residues in "residual structure" (i.e.neither helical nor sheet structure), a four-residue helical segment, and a single residue mediating the transition into strand 4. Subsequent to strand 4, the polypeptide chain continues towards the central tunnel in the shape of  the "connecting segment", to form the innermost strand of the following b-sheet.

The angle by which the b-sheets are rotated around the central axis in a propeller-like molecule is 360/n , with n= number of symmetric elements.In tachylectin-2, the resulting 72 degree-angle would account for large spatial gaps between adjacent b-sheets, were it not for the large loop between strand 3 and strand 4, the connecting segment, and the presence of large side chains in strand 4 (the latter is a common feature in molecules adopting a b-propeller structure).
The rather hydrophilic core structure of b-propellers sets this large and diverse group apart from most other proteins which are generally assumed to be built around a hydrophobic core, with more hydrophilic structural elements on the surface.  Tachylectin-2 is distinct from other b-propeller molecules in that a ring of conserved PHE residues lines the central tunnel. The electrophilic benzyl side chains of the PHE residues border the water molecules in the central tunnel which are configured in a specific dodecahedral arrangement that corresponds to the overall pentagonal symmetry of the tachylectin-2 molecule (Fig.3)
On the level of sequence elements resembling domains, five 47-residue tandem repeats with 49-68% sequence identity are prominent.They form important structural elements of the five ligand-binding pockets.(Image Fig.2)
 
 

A 2.1 Ligand-binding Site Topology
Due to its symmetry and the structural equivalence of each blade, tachylectin-2 provides 5 equivalent carbohydrate-binding sites.(Image Fig 1)
Each individual binding site is formed by structural elements contributed by two adjacent b sheets.The connecting segment following strand 4 of blade n-1 and the loop between strand 3 and 4 of blade n form much of the carbohydrate binding site. Although two different blades are coordinated upon ligand binding, the inherent rigidity of the polypeptide chain is such that binding of monomeric N-acetyl-glucosamine does not alter main conformation, and not even side chain conformation.(Table I; Image Fig. 5)
Beisel et al. (EMBO J. 18(9), 2313 - 2322, 1999 ) have analyzed and described the structural features of N-acetyl-glucosamine binding to the binding site in blade IV.(Image Fig. 6). 
The back portion of the binding site is formed by PHE137 and TRP169, while the sequence ASP167 to TRP169 forms the bottom portion.The right side of the binding pocket in Fig. 6 is bounded by the side chains of ASN168 and Leu170; the left side is defined by GLY10 to TRP134. The pocket environment changes considerably in polarity from the hydrophobic back portion to the rather hydrophilic left side which recognizes the two alcoholic HO-functions in 3 and 4-position of the hexose ring and the charged nitrogen of the acetamido function of the sugar side chain.What appears to be "unordered structure" in conventional assessment of secondary structure elements, is in fact a highly ordered network of polar bridges that serves to stabilize the ligand binding site. In blade IV, ASP167 forms a salt bridge with ARG172 that form a hydrogen bond with LEU160. the carbonyl atom of LEU160 participates in hydrogen bonds with two amino acids, namely ARG172 and TRP169. The ASN165 carbonyl oxygen forms a bond with the 4-OH function of the hexose ring; the ASP167 carbonyl oxygen forms a bond with the 3-OH function of the hexose ring. Additionally, the VAL129 carbonyl oxygen, together with ASP167, fixes a water molecule in place which forms a bond with the hexose 3-OH function.The acetamido methyl group is in van der Waals-contact with TRP134 and oriented into the hydrophobic pocket formed by the side chains of PHE137, TRP134, and LEU170. GLY36 is strictly conserved because any side chain in this position would "clog" the sugar binding site in each respective blade.
The engagement of the sugar 4-OH function in hydrogen bonding with two different amino acids explains the absence of significant affinity for carbohydrates bearing a bulky substitute in 4-position, such as N-acetyl-lactosamine. The difference in affinity towards the monomeric carbohydrate N-acetyl-glucosamine over N-acetyl-galactosamine can also be linked to the coordination of the 4-OH group: the axial position in N-acetyl-galactosamine is less favourable for interaction than the equatorial position in N-acetyl-glucosamine. In contrast, neither the hexose ring oxygen nor the 1-OH or 6-OH functions are engaged in coordination with amino acids.

A 2.0 Angstrom electron density map of the carbohydrate-binding site is shown in Image Fig. 4 

A stereo plot of the carbohydrate binding site within a blade is shown in Image Fig 7:
 

A3 Images Generated with RasMol

A4 Definitions of tachylectin-2 
with SCOP and CATH database algorithms 
A 4.1 SCOP: Structural Classification of Proteins
<http://scop.mrc-lmb.cam.ac.uk/scop/index.html> 
<http://scop.mrc-lmb.cam.ac.uk/scop/mail.cgi> 
<http://scop.mrc-lmb.cam.ac.uk/scop/help.html> 
<http://scop.mrc-lmb.cam.ac.uk/scop/data/scop.b.html> 
<http://scop.mrc-lmb.cam.ac.uk/scop/data/scop.b.c.html> 
<http://scop.mrc-lmb.cam.ac.uk/scop/data/scop.b.c.he.A.A.html> 

Fold: 5-bladed beta-propeller
consists of five 4-stranded beta-sheet motifs; meander

Lineage:
1. Root: http://scop.mrc-lmb.cam.ac.uk/scop/data/scop.b.html scop

2. Class: http://scop.mrc-lmb.cam.ac.uk/scop/data/scop.b.c.html 
            All beta proteins

3. Fold: http://scop.mrc-lmb.cam.ac.uk/scop/data/scop.b.c.he.A.A.A.html
           5-bladed beta-propeller
            consists of five 4-stranded beta-sheet motifs; meander

Superfamilies:
1. http://scop.mrc-lmb.cam.ac.uk/scop/data/scop.b.c.he.b.A.A.html 
    Tachylectin-2 (1)
2. http://scop.mrc-lmb.cam.ac.uk/scop/data/scop.b.c.he.b.b.A.html 
    Tachylectin-2 (1).,Tachylectin-2
3. http://scop.mrc-lmb.cam.ac.uk/scop/data/scop.b.c.he.b.b.b.html
    Japanese horseshoe crab (Tachypleus tridentatus) (1)
1. http://scop.mrc-lmb.cam.ac.uk/scop/pdb.cgi?sid=d1tl2a_ 1tl2. <http://www3.ncbi.nlm.nih.gov/htbin-post/Entrez/query?db=t&for;/=6&Dop;/=s&ui;;=1tl2> 
3. <http://scop.mrc-lmb.cam.ac.uk/scop/rsgen.cgi?pd=1tl2;pc=a> 
4. <http://scop.mrc-lmb.cam.ac.uk/scop/rsgen.cgi?chime=1;pd=1tl2;pc=a> 
1. <http://scop.mrc-lmb.cam.ac.uk/scop/pdb.cgi?sid=d1tl2a_>
2. chain a 
3. <http://scop.mrc-lmb.cam.ac.uk/scop/rsgen.cgi?pd=1tl2;pc=a> 
4. <http://scop.mrc-lmb.cam.ac.uk/scop/rsgen.cgi?chime=1;pd=1tl2;pc=a> 
5. <http://scop.mrc-lmb.cam.ac.uk/scop/search.cgi?sid=d1tl2a_&tle;/=px> 
Generated from scop database 1.57 with scopm 1.096

A 4.2 CATH

"http://www.biochem.ucl.ac.uk/bsm/cath_new/images/1fupA2.gif"
"http://www.biochem.ucl.ac.uk/bsm/cath_new/Impala/"
"http://www.biochem.ucl.ac.uk/bsm/cath_new/Gene3D/
"ftp://ftp.biochem.ucl.ac.uk/pub/cathdata/v2.4/" CLASS="TopMenu">

Domain 1tl2A0

Fold relatives

There are either no other non-identical relatives within this fold group or the structural comparisons for this domain have not yet been calculated.

Click here for the results of "structural neighbour"-analysis.
Click here for the results of ClustalW analysis.
 
 

B  Technical Section on Tachylectin Crystallography
B1 Materials and methods employed for Crystallography
Crystallization, data collection and derivatization
Crystallization was carried out in ChrysChem plates by the vapor diffusion method using 100 mM sodium acetate, pH 4.6, 2.0 M sodium formate as precipitant against 800 ml of precipitant in the reservoir; sitting drops were set up with 2.2 ml of protein solution and 1.5 ml of precipitation buffer. Within 2 days, crystals (Table II) appeared at 20 degrees Celsius up to a final size of ~600x200x150 mm³.They belonged to the trigonal space group P3₁21, having unit cell dimensions a=b= 89.42 Angstrom, c=73.38 Angstrom (NATI1), one molecule per asymmetric unit and V_M= 3.14 Angstrom³/Da corresponding to a 61% solvent content (Matthews B.W., J. Mol. Biol. 33, 491 - 497, 1968).
To prepare tachylectin-2 heavy atom derivatives, the crystals were soaked in a solution of the heavy atom compound in freshly prepasred precipitation buffer. Harvesting the crystals in fresh precipitant solution changed the unit cell dimensions to a=b=90.61 Angstrom, c=71.43 Angstrom (Table II).Therefore, a second data set (NATI2) was measured in order to analyze the heavy atom derivative diffraction data. Interpretable results were obtained with Pb(AcO)₂ (10 mM, 10 days), Me₃PbCl (10 mg/ml, 2 days) and the double derivative Pb(AcO)₂/Me₃PbCl (5 mg/ml each, 4 days).
The complex of tachylectin-2 with GlcNAc was prepared by cocrystallizing the protein using the conditions described above, but with 5 mM GlcNAc (Sigma) added to the precipitation buffer.These crystals belong to the same crystal form as the native ones (Table II).
All diffraction data were collected using a 300 mm MAR Research (Hamburg, Germany) image plate detector mounted on a Rigaku (Tokyo, Japan) RU200 rotating anode X-ray generator with graphite monochromatized CuK_a radiation. All image plate data were processed with MOSFLM (Leslie AGW, In: Moras D, Podjarny AD, Thierry JC (eds) Crystallographic Computing 5, Oxford University Press, Oxford 1991, pp. 50 - 61) and the CCP4 program suite (Collaborative Computational Project, Number 4, 1994).

Phase calculation, model building and refinement
The structure of tachylectin-2 was solved by the multiple isomorphous replacement (MIR) method using the three heavy atom derivatives described above. All derivative data were analyzed with the native data set NATI2, first using isomorphous Patterson maps. Heavy atom locations were confirmed by difference Fourier methods with appropriate initial single isomorphous replacement phases using CCP4 programs, as well as by Paterson vector superposition methods implemented in SHELX097 (Sheldrick G,  In: Moras D, Podjarny AD, Thierry JC (eds.) Crystallographic Computing 5, Oxford University Press, Oxford 1991, pp.145 - 157).The refinement of heavy atom parameters and calculation of MIR phases were done with SHARP (La Fortelle and Bricogne, Meth. Enzymol 276, 472 - 494, 1997). The final parameters are given in Table III. The initial MIR phases were improved with SOLOMON (Abrahams and Leslie, Acta Crystallogr. D52, 30 - 42), resulting in a 2.2 Angstrom electron density map that was interpretable in terms of a protein structure. All model building was done with FRODO (Jones TA, J. Appl. Crystallogr. 11, 268 - 272, 1978). Refinement was performed by conjugate gradient and simulated annealing protocols as implemented in X-PLOR 3.851 (Bruenger A, Kuriyan J, Karplus M, Science 235, 458 - 460, 1987).The final cycle of refinement was performed with REFMAC (Murshudav GM, Vagin AA, Dodson EJ, Acta Crystallogr. D53, 240 - 255, 1997) using the conjugate direction method with a maximum likelihood residual and a bulk solvent model. All protocols included refinement of individual isotropic B-factors. In order to use the higher resolution (2.0 Angstrom) data set NATI1 for refinement, the initial model built in the NATI2 cell was placed with X-PLOR rigid body refinement in the NATI1 cell, and used for all subsequent refinement cycles of tachylectin-2. The initial R-factor for the first refinement cycle of the unrefined model including all side chains was 31% (R_free = 35.6%). It dropped to 18.6% (R_free  = 22.9%, resolution range 15.0 to 2.0 Angstrom) for the final model including 129 water molecules. The water model was calculated using ARP ( Lamzin VS and Wilson KS, Acta Crystallograph. D49, 129 - 147) and verified by visual inspection. The final refinement statistics are shown in Table IV.
The X-ray structure of tachylectin-2 in complex with GlcNAc was solved using the model of the native protein excluding water molecules and calculating 2mF₀- DF_c and mF₀- DF_c
style electron density maps after five cycles of refinement with REFMAC. After adding the sugar molecules, the model was refined further by X-PLOR and REFMAC using the same protocols as described above. The water model was calculated and refined with ARP. The R-factor of the final model including 158 water molecules was 16.2% (R_free  = 20.2%, resolution range 15.0 - 2.0 Angstrom). The final refinement statistics are shown in Table IV. In a Ramachandran plot of tachylectin-2 in complex with GlcNAC calculated with PROCHECK (Laskowski RA, MacArthur MW, Moss DW, Thornton JM, J. Appl. Crystallograph. 26, 283 - 291, 1993), 91.1% of the residues lie in the most favoured regions, 8.9% in additionally allowed regions.

B2  Structural Classification of Proteins: PDB entry 1TL2

HEADER SUGAR BINDING PROTEIN 14-DEC-98 1TL2
TITLE TACHYLECTIN-2 FROM TACHYPLEUS TRIDENTATUS (JAPANESE
HORSESHOE CRAB)
COMPND MOL_ID: 1;
MOLECULE: TACHYLECTIN-2;
CHAIN: A;
SYNONYM: L10;
OTHER_DETAILS: FORMERLY NAMED L10, NOW TACHYLECTIN-2
SOURCE MOL_ID: 1;
ORGANISM_SCIENTIFIC: TACHYPLEUS TRIDENTATUS;
ORGANISM_COMMON: JAPANESE HORSESHOE CRAB
KEYWDS ANIMAL LECTIN, HORSESHOE CRAB, N-ACETYLGLUCOSAMINE, BETA-
PROPELLER, SUGAR BINDING PROTEIN
EXPDTA X-RAY DIFFRACTION
AUTHOR H.-G.BEISEL,S.KAWABATA,S.IWANAGA,R.HUBER,W.BODE
REVDAT 1 15-DEC-99 1TL2 0
JRNL AUTH H.G.BEISEL,S.KAWABATA,S.IWANAGA,R.HUBER,W.BODE
JRNL TITL TACHYLECTIN-2: CRYSTAL STRUCTURE OF A SPECIFIC
JRNL TITL 2 GLCNAC/GALNAC-BINDING LECTIN INVOLVED IN THE
JRNL TITL 3 INNATE IMMUNITY HOST DEFENSE OF THE JAPANESE
JRNL TITL 4 HORSESHOE CRAB TACHYPLEUS TRIDENTATUS
JRNL REF EMBO J. V. 18 2313 1999
JRNL REFN ASTM EMJODG UK ISSN 0261-4189
REMARK 1

REMARK 2
RESOLUTION. 2.00 ANGSTROMS.

REMARK 3
REFINEMENT.
PROGRAM : REFMAC
AUTHORS : MURSHUDOV,VAGIN,DODSON
DATA USED IN REFINEMENT.
RESOLUTION RANGE HIGH (ANGSTROMS) : 2.00
RESOLUTION RANGE LOW (ANGSTROMS) : 15.0
DATA CUTOFF (SIGMA(F)) : 0.000
COMPLETENESS FOR RANGE (%) : 94.0
NUMBER OF REFLECTIONS : 20892

FIT TO DATA USED IN REFINEMENT.
CROSS-VALIDATION METHOD : THROUGHOUT
FREE R VALUE TEST SET SELECTION : RANDOM
R VALUE (WORKING + TEST SET) : NULL
R VALUE (WORKING SET) : 0.162
FREE R VALUE : 0.202
FREE R VALUE TEST SET SIZE (%) : 10.000
FREE R VALUE TEST SET COUNT : 2072

NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT.
PROTEIN ATOMS : 1905
NUCLEIC ACID ATOMS : 0
HETEROGEN ATOMS : 75
SOLVENT ATOMS : 158

B VALUES.
FROM WILSON PLOT (A**2) : NULL
MEAN B VALUE (OVERALL, A**2) : 27.80
OVERALL ANISOTROPIC B VALUE.
B11 (A**2) : NULL
B22 (A**2) : NULL
B33 (A**2) : NULL
B12 (A**2) : NULL
B13 (A**2) : NULL
B23 (A**2) : NULL

ESTIMATED OVERALL COORDINATE ERROR.
ESU BASED ON R VALUE (A): NULL
ESU BASED ON FREE R VALUE (A): NULL
ESU BASED ON MAXIMUM LIKELIHOOD (A): NULL
ESU FOR B VALUES BASED ON MAXIMUM LIKELIHOOD (A**2): NULL

RMS DEVIATIONS FROM IDEAL VALUES.
DISTANCE RESTRAINTS. RMS SIGMA
BOND LENGTH (A) : 0.009 ; NULL
ANGLE DISTANCE (A) : 0.025 ; NULL
INTRAPLANAR 1-4 DISTANCE (A) : NULL ; NULL
H-BOND OR METAL COORDINATION (A) : NULL ; NULL

PLANE RESTRAINT (A) : NULL ; NULL
CHIRAL-CENTER RESTRAINT (A**3) : NULL ; NULL

NON-BONDED CONTACT RESTRAINTS.
SINGLE TORSION (A) : NULL ; NULL
MULTIPLE TORSION (A) : NULL ; NULL
H-BOND (X...Y) (A) : NULL ; NULL
H-BOND (X-H...Y) (A) : NULL ; NULL

CONFORMATIONAL TORSION ANGLE RESTRAINTS.
SPECIFIED (DEGREES) : NULL ; NULL
PLANAR (DEGREES) : NULL ; NULL
STAGGERED (DEGREES) : NULL ; NULL
TRANSVERSE (DEGREES) : NULL ; NULL

ISOTROPIC THERMAL FACTOR RESTRAINTS. RMS SIGMA
MAIN-CHAIN BOND (A**2) : NULL ; NULL
MAIN-CHAIN ANGLE (A**2) : NULL ; NULL
SIDE-CHAIN BOND (A**2) : NULL ; NULL
SIDE-CHAIN ANGLE (A**2) : NULL ; NULL

OTHER REFINEMENT REMARKS: NULL

REMARK 4
1TL2 COMPLIES WITH FORMAT V. 2.3, 09-JULY-1998

REMARK 100
THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 15-DEC-1998.
THE RCSB ID CODE IS RCSB000279.

REMARK 200
EXPERIMENTAL DETAILS
EXPERIMENT TYPE : X-RAY DIFFRACTION
DATE OF DATA COLLECTION : NULL
TEMPERATURE (KELVIN) : 289.0
PH : 4.60
NUMBER OF CRYSTALS USED : 1

SYNCHROTRON (Y/N) : N
RADIATION SOURCE : NULL
BEAMLINE : NULL
X-RAY GENERATOR MODEL : NULL
MONOCHROMATIC OR LAUE (M/L) : M
WAVELENGTH OR RANGE (A) : 1.5418
MONOCHROMATOR : NULL
OPTICS : NULL

DETECTOR TYPE : IMAGE PLATE
DETECTOR MANUFACTURER : MARRESEARCH
INTENSITY-INTEGRATION SOFTWARE : MOSFLM
DATA SCALING SOFTWARE : SCALA

NUMBER OF UNIQUE REFLECTIONS : 20896
RESOLUTION RANGE HIGH (A) : 2.000
RESOLUTION RANGE LOW (A) : 15.200
REJECTION CRITERIA (SIGMA(I)) : 2.000

OVERALL.
COMPLETENESS FOR RANGE (%) : 93.7
DATA REDUNDANCY : 4.000
R MERGE (I) : 0.07400
R SYM (I) : NULL
FOR THE DATA SET : NULL

IN THE HIGHEST RESOLUTION SHELL.
HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 2.03
HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 2.43
COMPLETENESS FOR SHELL (%) : 92.2
DATA REDUNDANCY IN SHELL : NULL
R MERGE FOR SHELL (I) : 0.17100
R SYM FOR SHELL (I) : NULL
FOR SHELL : NULL

DIFFRACTION PROTOCOL: SINGLE WAVELENGTH
METHOD USED TO DETERMINE THE STRUCTURE: MIR
SOFTWARE USED: CCP4, SHELXS-97, SHARP
STARTING MODEL: NULL

REMARK: NULL

CRYSTAL
SOLVENT CONTENT, VS (%): 61.0
MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 3.14

CRYSTALLIZATION CONDITIONS: NULL

REMARK 290
CRYSTALLOGRAPHIC SYMMETRY
SYMMETRY OPERATORS FOR SPACE GROUP: P 31 2 1

SYMOP SYMMETRY
NNNMMM OPERATOR
1555 X,Y,Z
2555 -Y,X-Y,1/3+Z
3555 -X+Y,-X,2/3+Z
4555 Y,X,-Z
5555 X-Y,-Y,2/3-Z
6555 -X,-X+Y,1/3-Z

WHERE NNN -> OPERATOR NUMBER
MMM -> TRANSLATION VECTOR

CRYSTALLOGRAPHIC SYMMETRY TRANSFORMATIONS
THE FOLLOWING TRANSFORMATIONS OPERATE ON THE ATOM/HETATM
RECORDS IN THIS ENTRY TO PRODUCE CRYSTALLOGRAPHICALLY
RELATED MOLECULES.
SMTRY1 1 1.000000 0.000000 0.000000 0.00000
SMTRY2 1 0.000000 1.000000 0.000000 0.00000
SMTRY3 1 0.000000 0.000000 1.000000 0.00000
SMTRY1 2 -0.500000 -0.866025 0.000000 0.00000
SMTRY2 2 0.866025 -0.500000 0.000000 0.00000
SMTRY3 2 0.000000 0.000000 1.000000 24.54000
SMTRY1 3 -0.500000 0.866025 0.000000 0.00000
SMTRY2 3 -0.866025 -0.500000 0.000000 0.00000
SMTRY3 3 0.000000 0.000000 1.000000 49.08000
SMTRY1 4 -0.500000 0.866025 0.000000 0.00000
SMTRY2 4 0.866025 0.500000 0.000000 0.00000
SMTRY3 4 0.000000 0.000000 -1.000000 0.00000
SMTRY1 5 1.000000 0.000000 0.000000 0.00000
SMTRY2 5 0.000000 -1.000000 0.000000 0.00000
SMTRY3 5 0.000000 0.000000 -1.000000 49.08000
SMTRY1 6 -0.500000 -0.866025 0.000000 0.00000
SMTRY2 6 -0.866025 0.500000 0.000000 0.00000
SMTRY3 6 0.000000 0.000000 -1.000000 24.54000

REMARK: NULL

REMARK 300
BIOMOLECULE: 1
THIS ENTRY CONTAINS THE CRYSTALLOGRAPHIC ASYMMETRIC UNIT
WHICH CONSISTS OF 1 CHAIN(S). SEE REMARK 350 FOR
INFORMATION ON GENERATING THE BIOLOGICAL MOLECULE(S).
 

REMARK 350
GENERATING THE BIOMOLECULE
COORDINATES FOR A COMPLETE MULTIMER REPRESENTING THE KNOWN
BIOLOGICALLY SIGNIFICANT OLIGOMERIZATION STATE OF THE
MOLECULE CAN BE GENERATED BY APPLYING BIOMT TRANSFORMATIONS
GIVEN BELOW. BOTH NON-CRYSTALLOGRAPHIC AND
CRYSTALLOGRAPHIC OPERATIONS ARE GIVEN.

BIOMOLECULE: 1
APPLY THE FOLLOWING TO CHAINS: A
BIOMT1 1 1.000000 0.000000 0.000000 0.00000
BIOMT2 1 0.000000 1.000000 0.000000 0.00000
BIOMT3 1 0.000000 0.000000 1.000000 0.00000

MISSING RESIDUES
THE FOLLOWING RESIDUES WERE NOT LOCATED IN THE
EXPERIMENT. (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN
IDENTIFIER; SSSEQ=SEQUENCE NUMBER; I=INSERTION CODE.)

M RES C SSSEQ I
VAL A 1
REMARK 500
GEOMETRY AND STEREOCHEMISTRY
SUBTOPIC: COVALENT BOND ANGLES

THE STEREOCHEMICAL PARAMETERS OF THE FOLLOWING RESIDUES
HAVE VALUES WHICH DEVIATE FROM EXPECTED VALUES BY MORE
THAN 6*RMSD (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN
IDENTIFIER; SSEQ=SEQUENCE NUMBER; I=INSERTION CODE).

STANDARD TABLE:

FORMAT: (10X,I3,1X,A3,1X,A1,I4,A1,3(1X,A4,2X),12X,F5.1)

EXPECTED VALUES: ENGH AND HUBER, 1991

M RES CSSEQI ATM1 ATM2 ATM3
PHE A 236 CA - C - O ANGL. DEV. =-15.4 DEGREES
DBREF 1TL2 A 1 236 GB 1256941 D45909 20 255
SEQADV 1TL2 VAL A 129 GB 1256941 ILE 148 CONFLICT
SEQADV 1TL2 TYR A 213 GB 1256941 HIS 232 CONFLICT
SEQRES 1 A 236 VAL GLY GLY GLU SER MET LEU ARG GLY VAL TYR GLN ASP
SEQRES 2 A 236 LYS PHE TYR GLN GLY THR TYR PRO GLN ASN LYS ASN ASP
SEQRES 3 A 236 ASN TRP LEU ALA ARG ALA THR LEU ILE GLY LYS GLY GLY
SEQRES 4 A 236 TRP SER ASN PHE LYS PHE LEU PHE LEU SER PRO GLY GLY
SEQRES 5 A 236 GLU LEU TYR GLY VAL LEU ASN ASP LYS ILE TYR LYS GLY
SEQRES 6 A 236 THR PRO PRO THR HIS ASP ASN ASP ASN TRP MET GLY ARG
SEQRES 7 A 236 ALA LYS LYS ILE GLY ASN GLY GLY TRP ASN GLN PHE GLN
SEQRES 8 A 236 PHE LEU PHE PHE ASP PRO ASN GLY TYR LEU TYR ALA VAL
SEQRES 9 A 236 SER LYS ASP LYS LEU TYR LYS ALA SER PRO PRO GLN SER
SEQRES 10 A 236 ASP THR ASP ASN TRP ILE ALA ARG ALA THR GLU VAL GLY
SEQRES 11 A 236 SER GLY GLY TRP SER GLY PHE LYS PHE LEU PHE PHE HIS
SEQRES 12 A 236 PRO ASN GLY TYR LEU TYR ALA VAL HIS GLY GLN GLN PHE
SEQRES 13 A 236 TYR LYS ALA LEU PRO PRO VAL SER ASN GLN ASP ASN TRP
SEQRES 14 A 236 LEU ALA ARG ALA THR LYS ILE GLY GLN GLY GLY TRP ASP
SEQRES 15 A 236 THR PHE LYS PHE LEU PHE PHE SER SER VAL GLY THR LEU
SEQRES 16 A 236 PHE GLY VAL GLN GLY GLY LYS PHE TYR GLU ASP TYR PRO
SEQRES 17 A 236 PRO SER TYR ALA TYR ASP ASN TRP LEU ALA ARG ALA LYS
SEQRES 18 A 236 LEU ILE GLY ASN GLY GLY TRP ASP ASP PHE ARG PHE LEU
SEQRES 19 A 236 PHE PHE
HET NAG 237 15
HET NAG 238 15
HET NAG 239 15
HET NAG 240 15
HET NAG 241 15
HETNAM NAG N-ACETYL-D-GLUCOSAMINE
HETSYN NAG NAG
FORMUL 2 NAG 5(C8 H15 N1 O6)
FORMUL 7 HOH *158(H2 O1)
HELIX 1 1 TRP A 28 ARG A 31 1 4
HELIX 2 2 TRP A 75 ARG A 78 1 4
HELIX 3 3 TRP A 87 GLN A 89 5 3
HELIX 4 4 TRP A 122 ARG A 125 1 4
HELIX 5 5 TRP A 134 GLY A 136 5 3
HELIX 6 6 TRP A 169 ARG A 172 1 4
HELIX 7 7 TRP A 181 THR A 183 5 3
HELIX 8 8 TRP A 216 ARG A 219 1 4
HELIX 9 9 TRP A 228 ASP A 230 5 3
SHEET 1 A 4 PHE A 231 PHE A 235 0
SHEET 2 A 4 LEU A 7 TYR A 11 -1 N VAL A 10 O ARG A 232
SHEET 3 A 4 LYS A 14 GLY A 18 -1 N GLY A 18 O LEU A 7
SHEET 4 A 4 THR A 33 GLY A 36 -1 N GLY A 36 O PHE A 15
SHEET 1 B 4 PHE A 45 LEU A 48 0
SHEET 2 B 4 GLU A 53 LEU A 58 -1 N VAL A 57 O PHE A 45
SHEET 3 B 4 LYS A 61 THR A 66 -1 N GLY A 65 O LEU A 54
SHEET 4 B 4 LYS A 80 GLY A 83 -1 N GLY A 83 O ILE A 62
SHEET 1 C 4 PHE A 92 PHE A 95 0
SHEET 2 C 4 TYR A 100 SER A 105 -1 N VAL A 104 O PHE A 92
SHEET 3 C 4 LYS A 108 SER A 113 -1 N ALA A 112 O LEU A 101
SHEET 4 C 4 THR A 127 GLY A 130 -1 N GLY A 130 O LEU A 109
SHEET 1 D 4 PHE A 137 PHE A 142 0
SHEET 2 D 4 TYR A 147 HIS A 152 -1 N VAL A 151 O LYS A 138
SHEET 3 D 4 GLN A 155 LEU A 160 -1 N ALA A 159 O LEU A 148
SHEET 4 D 4 THR A 174 GLY A 177 -1 N GLY A 177 O PHE A 156
SHEET 1 E 4 PHE A 184 PHE A 189 0
SHEET 2 E 4 THR A 194 GLN A 199 -1 N VAL A 198 O LYS A 185
SHEET 3 E 4 LYS A 202 TYR A 207 -1 N ASP A 206 O LEU A 195
SHEET 4 E 4 LYS A 221 ASN A 225 -1 N GLY A 224 O PHE A 203
CRYST1 89.460 89.460 73.620 90.00 90.00 120.00 P 31 2 1 6
ORIGX1 1.000000 0.000000 0.000000 0.00000
ORIGX2 0.000000 1.000000 0.000000 0.00000
ORIGX3 0.000000 0.000000 1.000000 0.00000
SCALE1 0.011178 0.006454 0.000000 0.00000
SCALE2 0.000000 0.012907 0.000000 0.00000
SCALE3 0.000000 0.000000 0.013583 0.00000