TrypanoSome calmodulin binding proteins Detection, sequence patterns and expression
Trypanosomes undergo biochemical changes during differentiation critical for survival in their invertebrate and vertebrate hosts. Calmodulin (CaM), a multifunctional Ca$\sp{2+}$-binding protein, is altered in concentration and compartmentalization during differentiation, inferring that trypanosomes...
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| Format: | Thesis Electronic Book |
| Language: | English |
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1990
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| LEADER | 03817ntm a22003133 4500 | ||
|---|---|---|---|
| 001 | b10781db-c6f0-4896-8cbc-192d794a93e8 | ||
| 005 | 20240901000000.0 | ||
| 008 | 110415s1990 xx |||||om||||||| ||eng d | ||
| 035 | |a (UMI)AAI9121119 | ||
| 035 | |a AAI9121119 | ||
| 040 | |a UMI |c UMI | ||
| 100 | 1 | |a Schleck, Sarah Ann | |
| 245 | 1 | 0 | |a TrypanoSome calmodulin binding proteins |h [electronic resource] : |b Detection, sequence patterns and expression |
| 260 | |c 1990 | ||
| 300 | |a 1 online resource (237 p.) | ||
| 500 | |a Source: Dissertation Abstracts International, Volume: 52-03, Section: B, page: 1226 | ||
| 502 | |a Thesis (Ph.D.)--Yale University, 1990 | ||
| 506 | |a Access is restricted by licensing agreement | ||
| 520 | |a Trypanosomes undergo biochemical changes during differentiation critical for survival in their invertebrate and vertebrate hosts. Calmodulin (CaM), a multifunctional Ca$\sp{2+}$-binding protein, is altered in concentration and compartmentalization during differentiation, inferring that trypanosomes use Ca$\sp{2+}$ as a second messenger in signal transduction as is common in higher eukaryotes. Native Trypanosoma brucei rhodesiense CaM (nTCaM) and recombinant TCaM (rTCaM) are identical based on genomic and amino acid sequence homologies, antigenicity, Ca$\sp{2+}$-dependent electrophoretic mobility shifts, peptide maps, activation of cAMP-dependent phosphodiesterase, and UV absorption spectra. Biotinylated rTCaM (Bio-rTCaM) labels high (100-110, 116-120, 140, 160, 180-200kD) and low (20-30kD) molecular weight polypeptides on Western blots of whole cell and crude membrane fractions in the presence of Ca$\sp{2+}$. Bio-rTCaM also binds only in the presence of Ca$\sp{2+}$ to affinity purified TCaM-binding proteins (M$\sb{\rm r}$ = 34, 37, 39, 42, 48, 55, 60, 65, 68, and 72kD) isolated from cytosolic fraction of bloodstream trypomastigotes on Western blots, of which p60, p65, p68, and p72 appear to be developmentally regulated. Antibodies against the CaM-binding proteins adducin, clathrin, MAP/tau and CaM-PKII and against a leishmanial paraflagellar rod associated protein cross react with peptides in bloodstream trypomastigote fractions | ||
| 520 | |a cDNA clones expressing Ca$\sp{2+}$/TCaM-binding peptides were plaque purified from a $\lambda$gt11 T. cruzi bloodstream trypomastigote cDNA library using Ca$\sp{2+}$/Bio-rTCaM as a functional ligand probe. Clone $\lambda$TcCaMBP13C, which contains a 542bp EcoRI insert with an open reading frame for an $\approx$15-20kD peptide, expresses a 135kD $\beta$-galactosidase fusion protein that also binds Ca$\sp{2+}$/Bio-rTCaM on Western blots. This clone and its gene product have been named CaMcruzin for T. cruzi TCaM-binding protein. The EcoRI cDNA insert hybridizes to one end of a 4.0kb SalI insert from a T. cruzi EMBL3 genomic clone based on Southern blots and digestion patterns. It also hybridizes to a 1.8-2.0kb band of epimastigote derived T. cruzi poly(A$\sp+$) mRNA that is maximally indicative of a 60-70kD protein. Ca$\sp{2+}$/Bio-rTCaM labels peptides of this size range on Western blots of T. cruzi epimastigote and trypomastigote homogenates. CaMcruzin is the first functionally identified and partially sequenced TCaM-binding protein reported for trypanosomes | ||
| 590 | |a Access is available to the Yale community | ||
| 650 | 4 | |a Biology, Microbiology | |
| 650 | 4 | |a Biology, Molecular | |
| 710 | 2 | |a Yale University | |
| 773 | 0 | |t Dissertation Abstracts International |g 52-03B | |
| 791 | |a Ph.D | ||
| 792 | |a 1990 | ||
| 999 | 1 | 0 | |i b10781db-c6f0-4896-8cbc-192d794a93e8 |l 9839493 |s US-CTY |m trypanosome_calmodulin_binding_proteinsdetection_sequence_patterns_andelect1990____________t________________________________________schleck__sarah_ann_________________e |
| 999 | 1 | 1 | |l 9839493 |s ISIL:US-CTY |t BKS |a yulintx |c None |p UNLOANABLE |