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PDBsum entry 1ex3

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protein links
Hydrolase PDB id
1ex3
Jmol
Contents
Protein chain
245 a.a. *
Waters ×42
* Residue conservation analysis
PDB id:
1ex3
Name: Hydrolase
Title: Crystal structure of bovine chymotrypsinogen a (tetragonal)
Structure: Chymotrypsinogen a. Chain: a. Ec: 3.4.21.1
Source: Bos taurus. Cattle. Organism_taxid: 9913. Organ: pancreas
Resolution:
3.00Å     R-factor:   0.204    
Authors: P.E.Pjura,A.M.Lenhoff,S.A.Leonard,A.G.Gittis
Key ref:
P.E.Pjura et al. (2000). Protein crystallization by design: chymotrypsinogen without precipitants. J Mol Biol, 300, 235-239. PubMed id: 10873462 DOI: 10.1006/jmbi.2000.3851
Date:
28-Apr-00     Release date:   17-May-00    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chain
Pfam   ArchSchema ?
P00766  (CTRA_BOVIN) -  Chymotrypsinogen A
Seq:
Struc:
245 a.a.
245 a.a.
Key:    PfamA domain  Secondary structure  CATH domain

 Enzyme reactions 
   Enzyme class: E.C.3.4.21.1  - Chymotrypsin.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: Preferential cleavage: Tyr-|-Xaa, Trp-|-Xaa, Phe-|-Xaa, Leu-|-Xaa.
 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     extracellular region   2 terms 
  Biological process     digestion   2 terms 
  Biochemical function     catalytic activity     6 terms  

 

 
DOI no: 10.1006/jmbi.2000.3851 J Mol Biol 300:235-239 (2000)
PubMed id: 10873462  
 
 
Protein crystallization by design: chymotrypsinogen without precipitants.
P.E.Pjura, A.M.Lenhoff, S.A.Leonard, A.G.Gittis.
 
  ABSTRACT  
 
Protein crystals are usually obtained by an empirical approach based on extensive screening to identify suitable crystallization conditions. In contrast, we have used a systematic predictive procedure to produce data-quality crystals of bovine chymotrypsinogen A and used them to obtain a refined X-ray structure to 3 A resolution. Measurements of the osmotic second virial coefficient of chymotrypsinogen solutions were used to identify suitable solvent conditions, following which crystals were grown for approximately 30 hours by ultracentrifugal crystallization, without the use of any precipitants. Existing structures of chymotrypsinogen were obtained in solutions including 10-30 % ethanol, whereas simple buffered NaCl solutions were used here. The protein crystallized in the tetragonal space group P4(1)2(1)2, with one molecule per asymmetric unit. The quality of the refined map was very high throughout, with the main-chain atoms of all but four residues clearly defined and with nearly all side-chains also defined. Although only minor differences are seen compared to the structures previously reported, they indicate the possibility of structural changes due to the crystallization conditions used in those studies. Our results show that more systematic crystallization of proteins is possible, and that the procedure can expand the range of conditions under which crystals can be grown successfully and can make new crystal forms available.
 
  Selected figure(s)  
 
Figure 1.
Figure 1. Values of the osmotic second virial coefficient of chymotrypsinogen as a function of solution composition, measured by membrane osmometry (open symbols; this work) and static light scattering (filled symbols) [Velev et al 1998]. Osmotic pressure measurements were made of solutions of a-chymotrypsinogen A (Sigma, Type II, from bovine pancreas; 6× crystallized and lyophilized powder, used without further purification). The six solvents each contained 0.1 or 0.3 M NaCl, 10 mM citric acid, and 0.05 % Na azide, and were adjusted to final pH with NaOH or HCl as required. Samples were filtered through 0.22 µm filters before use. Protein concentrations were determined by measurement of A[280]. Measurements were made at 25°C on a Knauer A0330 membrane osmometer using vacuum-degassed Millipore PLBC 3000 MWCO regenerated cellulose ultrafiltration membranes. Measurements were made at six different protein concentrations, ranging from 2 to 12 g/l, for each solvent and in triplicate for each concentration. The data were least-squares fitted to equation (1) (truncated) to extract the number-average molecular mass and B[22].
Figure 2.
Figure 2. View of monomer packing in the unit cell of the new crystal form of chymotrypsinogen, showing the large solvent channels penetrating the unit cell.
 
  The above figures are reprinted by permission from Elsevier: J Mol Biol (2000, 300, 235-239) copyright 2000.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
19720033 R.Høiberg-Nielsen, P.Westh, L.K.Skov, and L.Arleth (2009).
Interrelationship of steric stabilization and self-crowding of a glycosylated protein.
  Biophys J, 97, 1445-1453.  
17766383 A.C.Dumetz, A.M.Snellinger-O'brien, E.W.Kaler, and A.M.Lenhoff (2007).
Patterns of protein protein interactions in salt solutions and implications for protein crystallization.
  Protein Sci, 16, 1867-1877.  
17704182 W.F.Weiss, T.K.Hodgdon, E.W.Kaler, A.M.Lenhoff, and C.J.Roberts (2007).
Nonnative protein polymers: structure, morphology, and relation to nucleation and growth.
  Biophys J, 93, 4392-4403.  
15090552 A.E.Schmidt, T.Ogawa, D.Gailani, and S.P.Bajaj (2004).
Structural role of Gly(193) in serine proteases: investigations of a G555E (GLY193 in chymotrypsin) mutant of blood coagulation factor XI.
  J Biol Chem, 279, 29485-29492.  
15034935 E.Pechkova, and C.Nicolini (2004).
Atomic structure of a CK2alpha human kinase by microfocus diffraction of extra-small microcrystals grown with nanobiofilm template.
  J Cell Biochem, 91, 1010-1020.  
12717013 E.Y.Chi, S.Krishnan, B.S.Kendrick, B.S.Chang, J.F.Carpenter, and T.W.Randolph (2003).
Roles of conformational stability and colloidal stability in the aggregation of recombinant human granulocyte colony-stimulating factor.
  Protein Sci, 12, 903-913.  
14580581 P.M.Tessier, and A.M.Lenhoff (2003).
Measurements of protein self-association as a guide to crystallization.
  Curr Opin Biotechnol, 14, 512-516.  
12486724 P.M.Tessier, H.R.Johnson, R.Pazhianur, B.W.Berger, J.L.Prentice, B.J.Bahnson, S.I.Sandler, and A.M.Lenhoff (2003).
Predictive crystallization of ribonuclease A via rapid screening of osmotic second virial coefficients.
  Proteins, 50, 303-311.  
11867474 P.M.Tessier, A.M.Lenhoff, and S.I.Sandler (2002).
Rapid measurement of protein osmotic second virial coefficients by self-interaction chromatography.
  Biophys J, 82, 1620-1631.  
11470603 P.R.Mittl, and M.G.Grütter (2001).
Structural genomics: opportunities and challenges.
  Curr Opin Chem Biol, 5, 402-408.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time.