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Biomedical subjects

C H Lo

Publications and source records attributed to C H Lo.

At least 19 recordsLinked to original sources
PubMed1997

Chemical selection for catalysis in combinatorial antibody libraries.

For the past decade the immune system has been exploited as a rich source of de novo catalysts. Catalytic antibodies have been shown to have chemoselectivity, enantioselectivity, large rate accelerations, and even an ability to reroute chemical reactions. In many instances catalysts have been made for reactions for which there are no known natural or man-made enzymes. Yet, the full power of this combinatorial system can only be exploited if there was a system that allows for the direct selection of a particular function. A method that allows for the direct chemical selection for catalysis from antibody libraries was so devised, whereby the positive aspects of hybridoma technology were preserved and re-formatted in the filamentous phage system to allow direct selection of catalysis. This methodology is based on a purely chemical selection process, making it more general than biologically based selection systems because it is not limited to reaction products that perturb cellular machinery.

Animals↗
PubMed1995

Reactive immunization.

For almost 200 years inert antigens have been used for initiating the process of immunization. A procedure is now described in which the antigen used is so highly reactive that a chemical reaction occurs in the antibody combining site during immunization. An organophosphorus diester hapten was used to illustrate this concept coined "reactive immunization." The organophosphonate recruited chemical potential from the immune response that resembled the way these compounds recruit the catalytic power of the serine hydrolases. During this recruitment, a large proportion of the isolated antibodies catalyzed the formation and cleavage of phosphonylated intermediates and subsequent ester hydrolysis. Reactive immunization can augment traditional immunization and enhance the scope of catalytic antibody chemistry. Among the compounds anticipated to be effective are those that contain appropriate reactive functionalities or those that are latently reactive, as in the mechanism-based inhibitors of enzymes.

Animals↗
PubMed1994

Direct selection for a catalytic mechanism from combinatorial antibody libraries.

Semisynthetic combinatorial antibody library methodology in the phage-display format was used to select for a cysteine residue in complementarity-determining regions. Libraries were panned with an alpha-phenethyl pyridyl disulfide that undergoes disulfide interchange. Out of 10 randomly picked clones, two contained an unpaired cysteine, one of which was studied. The antibody catalyzed the hydrolysis of the corresponding thioester where the electrophilic carbonyl occupies the three-dimensional space that was defined by the reactive sulfur atom during selection. The reaction operates by covalent catalysis. Although the steady-state rate enhancement relative to the activated thiol ester substrate is modest, hydrolysis of the acylated cysteine intermediate is remarkably efficient with a catalytic advantage of about four orders of magnitude. The results suggest that iterative mechanism-based selection procedures can recapitulate the enzymatic mechanisms refined through evolution.

Amino Acid Sequence↗
PubMed1991

Ostrich crystallins. Structural characterization of delta-crystallin with enzymic activity.

Lens crystallins from the African ostrich (Struthio camelus) were isolated and characterized. Four crystallin fractions corresponding to alpha-, delta/beta- and beta-crystallins similar to those of duck crystallins were isolated, but epsilon-crystallin was found to be absent. The native molecular masses and subunit structures of the purified fractions were analysed by gel filtration. SDS/PAGE and isoelectric focusing, revealing various extents of heterogeneity in each orthologous crystallin class. An ion-exchange chromatographic method was used for the large-scale preparation of delta-crystallin suitable for structural and enzymic studies. It was unexpectedly found that the purified native delta-crystallin of ostrich lens possessed high argininosuccinate lyase activity, in contrast with chicken delta-crystallin. The c.d. spectra indicated a predominant beta-sheet structure in alpha- and beta-crystallins, and a significant contribution of alpha-helical structure in the delta-crystallin fraction. The estimate of secondary structures from c.d. spectroscopy for each crystallin class bears a resemblance to that of duck crystallins, except that ostrich delta-crystallin possesses much less helical content than duck delta-crystallin. Comparison of crystallin compositions and structures from aquatic and terrestrial birds revealed distinct differences.

Amino Acids↗
PubMed1988

Biochemical comparison of lens crystallins from three reptilian species.

Lens crystallins were isolated from the homogenates of reptilian eye lenses derived from three different species by gel-permeation chromatography and characterized by gel electrophoresis, amino-acid analysis, N-terminal sequence analysis and circular dichroism. Four fractions corresponding to alpha-, delta/epsilon/beta-, beta- and gamma-crystallins were obtained for the crystallins from caiman lenses, whereas delta- and gamma-crystallin fraction were present in lesser amounts or missing in the turtle and snake lenses, respectively. The native molecular masses for these purified fractions and their polypeptide compositions were determined by gel filtration and SDS-gel electrophoresis, respectively, revealing the typical subunit compositions for each classified crystallin. The spectra of circular dichroism indicate a predominant beta-sheet structure in alpha-, beta- and gamma-crystallins, and a major contribution of alpha-helical structure in delta/epsilon-crystallin fraction, which bears a resemblance to the secondary structure of delta-crystallin from the chicken lenses. Comparison of the amino-acid contents of each orthologous class of reptilian crystallins with those of evolutionary distant species still exhibited similarity in their amino-acid compositions. N-terminal sequence analysis of the crystallin fractions revealed that all fractions except that of gamma-crystallin are N-terminally blocked. Extensive sequence similarity between the reptilian gamma-crystallin polypeptides and those from other vertebrate species were found, which establish the close relatedness of gamma-crystallins amongst the major classes of vertebrates.

Alligators and Crocodiles↗
PubMed1988

N-terminal sequences of gamma-crystallins from the amphibian lens and their homology with gamma-crystallins of other major classes of vertebrates.

gamma-Crystallins were isolated from the homogenate of frog eye lenses (Rana catesbeiana) by exclusion gel chromatography and further purified by cation-exchange chromatography. They were the only group of crystallins possessing free amino groups amenable to sequence analysis by Edman degradation. Comparison of the amino acid contents of the purified subfractions of gamma-crystallins indicated their close relatedness in amino acid compositions and probably sequence homology as well. The amino-terminal sequence analysis of the purified gamma-crystallin subfractions showed extensive homology between these amphibian gamma-crystallin polypeptides themselves and also those from other vertebrate species, suggesting the existence of a multigene family and their close relatedness to gamma-crystallins of other vertebrates. The sequence comparison of the gamma-crystallin polypeptides from all major classes of vertebrates has provided strong support for the divergent evolution of gamma-crystallin family.

Amino Acid Sequence↗
PubMed1987

Sequence comparison of gamma-crystallins from the reptilian and other vertebrate species.

Lens crystallins were isolated from homogenates of reptilian eye lenses (Caiman crocodylus apaporiensis) by gel-permeation chromatography and characterized by gel electrophoresis, and amino acid and N-terminal sequence analyses. Four fractions corresponding to alpha-, delta/epsilon/beta-, beta- and gamma-crystallins were identified on the basis of their electrophoretic patterns as revealed by SDS gel electrophoresis. Comparison of the amino acid contents of reptilian crystallins with those of mammals suggests that each orthologous class of crystallins from the evolutionarily distant species still exhibits similarity in their amino acid compositions and probably sequence homology as well. All fractions except that of gamma-crystallin were found to be N-terminally blocked. N-terminal sequence analysis of the purified gamma-crystallin subfractions showed extensive homology between the reptilian gamma-crystallin polypeptides themselves and also those from other vertebrate species, suggesting the existence of a multigene family and their close relatedness to gamma-crystallins of other vertebrates.

Alligators and Crocodiles↗
PubMed1979

Physical properties of human serum hexosaminidases A and B: studies in normal and cancer patients.

The major forms of human serum hexosaminidases A and B (Hex A and Hex B) were isolated from normal subjects and cancer patients using DEAE-cellulose. In normal serum, Hex A was heat-labile and had an apparent KM of 1.13 mM and Vmax of 0.51 mumol/ml/h; Hex B was heat-stable and had an apparent KM of 0.85 mM and Vmax of 0.22 mumol/ml/h. Both forms had the same pH optimum, at 4.3. Hex A and Hex B from the sera of cancer patients resembled their normal counterparts in heat stability, pH optimum, and apparent KM (1.07 mM for Hex A, 0.88 mM for Hex B). In contrast, the Vmax values for the cancer sera isozymes were greater than those of normal sera (0.70 and 0.40 mumol/ml/h for Hex A and Hex B, respectively).

Hexosaminidases↗
PubMed1978

Human serum hexosaminidase: elevated B form isozyme in cancer patients.

The activity of N-acetyl hexosaminidase (beta-2-acetamido-2-deoxy-D-glucoside:N-acetamido-deoxyglucohydrolase, E.C. 3.2.1.30) was significantly greater in sera of patients with solid malignant tumors than in sera of healthy volunteers or of patients with nonmalignant ailments. The increased activity was found in the two major isozymes, hexosaminidase A and hexosaminidase B, but the increase in the latter isozyme was more prominent than in the former. Measurement of the levels of the hexosaminidase isozymes may provide an ancillary diagnostic test for malignancy.

Adolescent↗