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Conjugated proteins

Conjugated proteins. Conjugated proteins = protein moiety + non protein part (prosthetic group) as carbohydrate, lipids, nucleic acid, etc..; linked together by covalent bond, or weak bonds (ionic, hydrogen, van der waals ).

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Conjugated proteins

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  1. Conjugated proteins Conjugated proteins = protein moiety + non protein part (prosthetic group) as carbohydrate, lipids, nucleic acid, etc..; linked together by covalent bond, or weak bonds (ionic, hydrogen, van derwaals). eg : hemoproteins, flavoproteins, glycoproteins, lipoproteins, phosphoproteins, nucleoproteins, metalloproteins, and proteoglycans.

  2. Conjugated proteins • Hemoproteins e.g:Hb , Myoglobin, cytochrome , catalase, peroxidase • It’s protein part + heme part ( tetra pyroleheterocycle bound to Fe+2) • Heme has peroxidase like reaction due to structure similarity between the prosthetic group of hemoglobin and peroxidase SH2 + H2O2 S + 2 H2O • Unlike peroxidase , heme retain its catalytic properties after boiling and treatment with strong acids (denaturating agents)

  3. HbexperImentPractical applications:very sensitive and used in trace analysis of blood for biochemical, medicolegal and clinical examinations

  4. 2. Phosphoproteins • E.g : casein (in milk), vitellin (in egg yolk), phosphorylase • Prosthetic group: H3PO4linked by ester bond to –OH of serine or threonine of protein moiety • Principle: Protein part tested by biuret reaction H3PO4 tested by amm. Molybdateamm. Phosphomolybdatereducing agent molybdenum blue ascorbic acid

  5. Nucloprotein • Abundant in nucleated cell such as liver, spleen • Not found in RBC’s • Classfied into RNP , DNP

  6. DNA isolation Spleen cells grind broken cells CTAB weakened cells Hypotonic saline spilled cytoplasm DNP Spilled cytoplasm = lipids+ proteins + nucleic acids + small molecule chloroform-isoamylalchol Organic phase (lipids) + Interphase (protein ppt) + Aqueous Phase (nucleic acid & small molecule). Aqueous phase Ethanol DNA (filaments) + RNA (gel ppt) + small molecule (in solution)

  7. DNA isolation CTAB detergant : solubilize cell membrane , inactivate nucleases EDTA ( chelating agent ) : cheleate necessary divalent and trivalent cationsfor nucleases activity so inhibit DNA degradation

  8. DNP identification • Protein part : biuretreaction (violet color) • H3PO4 part : molybdenum reaction (blue color) • Deoxy ribose : di-phenylamine test ( blue colour) • Purines: amm. AgNO3 test ( pale brown ppt)

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