The isolation and properties of the protein calliphorin.
1. A procedure for the isolation of the protein calliphorin from larvae and pupae of the blowfly Calliphora erythrocephala is described. 2. The calliphorin preparation shows a single component in the ultracentrifuge at pH6.3 and gives a single band when stained for protein after agar-gel or starch-gel electrophoresis at pH6.3 or 8.6. Immunoelectrophoresis yields only one arc, associated with the stained spot, to a rabbit antiserum known to react with 13 other soluble components of Calliphora pupae. 3. Calliphorin has s(0) (20,w) 19.4S, D(0) (20,w) 3.25x10(-7)cm(2).s(-1) and f/f(0) 1.22, indicating a molecular weight of 528000 and a compact symmetrical structure. The molecular weight determined by the meniscus-depletion sedimentation-equilibrium method is 529000. 4. In 6.2m-guanidine hydrochloride calliphorin dissociates into six components each with a molecular weight of about 87000. Calliphorin reversibly dissociates into components with sedimentation coefficients of about 7S as the pH is raised progressively above pH6.5. 5. Calliphorin has an unusually high tyrosine and phenylalanine content (442 and 400mol/mol of protein respectively), a relatively high methionine content (162mol/mol of protein) and very little cystine or cysteine (18mol/mol of protein). The E(280)/E(250) ratio is 3.2. The pure protein contains 0.4-0.5% carbohydrate. 6. When examined in the electron microscope by the negative staining technique the protein is seen to consist of particles which are right prisms, being 105A wide and 65A high, rectangular in side view and curvilinear equilateral triangles in surface view.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D003553:Cystine / Q000032:analysis; D004175:Diptera / Q000032:analysis*; D004586:Electrophoresis; D006146:Guanidines; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera; D007122:Immunoelectrophoresis; D007814:Larva; D008715:Methionine / Q000032:analysis; D008854:Microscopy, Electron; D008970:Molecular Weight; D010649:Phenylalanine / Q000032:analysis; D011506:Proteins / Q000302:isolation & purification*; D011679:Pupa; D014443:Tyrosine / Q000032:analysis; D014461:Ultracentrifugation
D000596:Amino Acids; D002241:Carbohydrates; D006146:Guanidines; D007106:Immune Sera; D011506:Proteins; D014443:Tyrosine; D010649:Phenylalanine; D003553:Cystine; D008715:Methionine