Anti-lysozyme antibody

Tello, D., E. Eisenstein, F.P. Schwarz, F.A. Goldbaum, B.A. Fields, R.A. Mariuzza, and R.J. Poljak. 1994. Structural and physicochemical analysis of the reaction between the anti-lysozyme antibody D1.3 and the anti-idiotopic antibodies E225 and E5.2. J Mol Recognit 7 57-62. 

In this exercise you will examine the interactions between the enzyme lysozyme (Chapter 4) and the Fab portion of the anti-lysozyme antibody. Use the PDB identifier 1FDL to explore the structure of the IgGl Fab fragment-lysozyme complex (antibody-antigen complex). View the structure using Protein Explorer, and also use the information in the PDBsum summary of the structure to answer the following questions. 

Hibbits, K. A., Gill, D. S., and Willson, R. C. (1994). Isothermal titration calorimetric study of the association of hen egg lysozyme and the anti-lysozyme antibody HyHEL-5. Biochemistry 33(12), 3584-3590. 

Tsumoto, K., K. Ogasahara, Y. Ueda, K. Watanabe, K. Yutani, and I. Kumagai. 1995. Role of Tyr residues in the contact region of anti-lysozyme monoclonal antibody HyHELlO for antigen binding. J Biol Chem 270 18551-18557. 

One of the 6 lysyl residues (residue 97) of lysozyme is near the active site. Conjugation of a hapten to this group does not affect the activity of the enzyme, but incubation of the enzyme-hapten conjugate with the specific anti-hapten antibody prior to the addition of the bulky substrate, produces serious inhibition. The smaller Fab is not as effective for inhibition as complete IgG. 

Enzyme activity can, therefore, be conveniently followed by measuring the formation of NADH at 340 nm. This enzyme is popular in AM-type EIA (Table 10.17). The reaction of anti-hapten antibodies with the hapten attached to the enzyme either inhibits or potentiates the enzyme activity, probably through changes in the conformation of the enzyme. MDase is detectable at 10 " M during a 1-min measurement. (Lysozyme is detectable at 10 M during the same time interval). 

Reduced dilutional and cleansing effect due to low secretion rate and low pH in saliva. Saliva and mucosa secretions have defense factors, such as lactoferrin, sialoperoxidase, lysozyme, histidine-rich polypeptide, secretory IgA antibodies, specific anti-Candida antibodies which help prevent adhesion and overgrowth of Candida. 

Recently, de la Paz et al. (242) modeled the binding sites of several antilysozyme monoclonal antibodies (mAbs) using both splicing and energy minimization techniques. Chothia and co-workers (88) used a similar approach to model their own anti-lysozyme mAb with more emphasis on preserving the orientation of the structural hypervariable residues. This model was recently confirmed by X-ray crystallography (243),... 

AltobelH, G., Subramaniam, S. Kinetics of association of anti-lysozyme monoclonal antibody D44.1 and hen-egg lysozyme. Biophys. J. 2000, 79,2954-65. 

The DNP-DAS label bound between two large protein molecules by means of noncovalent binding to an anti-TNP antibody on one side and on the other side via covalent linkage to lysozyme using cyanuric chloride as the cross-linker [10]. (Reproduced with permission from Elsevier.)... 

Lysozyme has been isolated from bovine and canine cartilages and from canine sera by a variety of chromatographic procedures, including affinity chromatography on carboxymethylchitin. The composition of the bovine lysozyme is similar to that of other mammalian lysozymes. Anti-(canine lysozyme) antibodies cross-reacted with bovine lysozyme, but with neither hen egg-white lysozyme nor embryonic chicken-cartilage lysozyme. Electron microscopy has shown that lysozyme in canine epiphyseal plates is localized extracellularly, mainly in the vicinity of the chondrocytes. Evidence that lysozyme has a physiological role in normal endochondral calcification has been presented. ... 

Goat antibodies specific for hen egg-white lysozyme, the loop-region (residues 60—83), and other regions reacted with the loop-region of the intact enzyme. Other egg-white lysozymes interfered in this reaction for example, quail lysozyme was as reactive as hen lysozyme in the anti-lysozyme system but it was much less reactive with anti-loop antibodies, whereas turkey lysozyme was as reactive as hen lysozyme with anti-loop antibodies but it differed in its reactivity with antilysozyme. Thus, it appears that the loop-region of hen egg-white lysozyme is the most reactive of all the loop-regions, and the results were discussed in terms of the amino-acid sequences of these lysozymes. 

To test DC-specific targeting, model antigens such as hen egg lysozyme (HEL) and ovalbumin (OVA) were covalently linked to anti-DEC-205 antibodies and injected into mice. This resulted in antigen loading of the lymph node DCs,... 

Kinetics of Anti-Hen Egg Lysozyme Monoclonal Antibodies Hyhel-5 and Hyhel-10. 

Slagle, S., R. E. Kozack and S. Subramaniam. (1994). Role of electrostatics in antibody-antigen association anti-hen egg lysozyme/lysozyme complex (HyHEL-5/HEL). J. Biomol. Struct. Dyn. 12 439-56. 

Specifically, an appropriately designed 2,4-dinitrophenyl-hapten derivative of fluorescent traws-4,4 -diaminostilbene (DAS) was squeezed between two large globular proteins lysozyme (Lys) from one side and anti-2,4,6-trinitrophenyl antibody (anti-TNP) from the other side, in order to provide the desired constricted environment to restrict traws/ds-stilbene isomerization within the anti-TN P-DNP-DAS-Lys adduct (Figure 10.13). 

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