Isolation and Sequence Comparison of a Maize Calmodulin cDNA

Griess, Eike A.; Igloi, Gabor L.; Feix, Günter

doi:10.1104/pp.104.4.1467

Cited by 6 publications

(6 citation statements)

References 1 publication

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“…Similar sequence differences in this amino acid position have previously been reported in cDNA-derived sequences for multiple isoforms of CaM present in a range of different higher plants (Table 1). Thus multiple isoforms of CaM that contain both sequences have been reported in corn, potato, and wheat (Jena et al, 1989;Griess et al, 1994;Toda et al, 1994;Breton et al, 1995;Takezawa et al, 1995;Yang et al, 1996), whereas cDNA-derived CaM isoforms that contain only Glu 7 (spinach) or Asp 7 (barley, leaf mustard, and red bryony) provide further support that there is a large amount of heterogeneity with respect to the identity of the acidic amino acid at this position encoded within the DNA sequences for plant CaM (Ling and Zielinski, 1989;Galaud et al, 1993;Chye et al, 1995). Our results provide strong support that two isoforms of CaM containing both sequences are expressed in wheat germ in approximately equal abundances.…”

Section: Identification Of Multiple Isoforms Of Cam Expressed In Wheat Germmentioning

confidence: 99%

Loss of Conformational Stability in Calmodulin upon Methionine Oxidation

Gao

Yin

Yao

et al. 1998

Biophysical Journal

162

169

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We have used electrospray ionization mass spectrometry (ESI-MS), circular dichroism (CD), and fluorescence spectroscopy to investigate the secondary and tertiary structural consequences that result from oxidative modification of methionine residues in wheat germ calmodulin (CaM), and prevent activation of the plasma membrane Ca-ATPase. Using ESI-MS, we have measured rates of modification and molecular mass distributions of oxidatively modified CaM species (CaMox) resulting from exposure to H2O2. From these rates, we find that oxidative modification of methionine to the corresponding methionine sulfoxide does not predispose CaM to further oxidative modification. These results indicate that methionine oxidation results in no large-scale alterations in the tertiary structure of CaMox, because the rates of oxidative modification of individual methionines are directly related to their solvent exposure. Likewise, CD measurements indicate that methionine oxidation results in little change in the apparent alpha-helical content at 28 degrees C, and only a small (0.3 +/- 0.1 kcal mol(-1)) decrease in thermal stability, suggesting the disruption of a limited number of specific noncovalent interactions. Fluorescence lifetime, anisotropy, and quenching measurements of N-(1-pyrenyl)-maleimide (PMal) covalently bound to Cys26 indicate local structural changes around PMal in the amino-terminal domain in response to oxidative modification of methionine residues in the carboxyl-terminal domain. Because the opposing globular domains remain spatially distant in both native and oxidatively modified CaM, the oxidative modification of methionines in the carboxyl-terminal domain are suggested to modify the conformation of the amino-terminal domain through alterations in the structural features involving the interdomain central helix. The structural basis for the linkage between oxidative modification and these global conformational changes is discussed in terms of possible alterations in specific noncovalent interactions that have previously been suggested to stabilize the central helix in CaM.

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Section: Identification Of Multiple Isoforms Of Cam Expressed In Wheat Germmentioning

confidence: 99%

Loss of Conformational Stability in Calmodulin upon Methionine Oxidation

Gao

Yin

Yao

et al. 1998

Biophysical Journal

162

169

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“…This result is similar to the analysis based on the P. yezoensis and Porphyra haitanensis ESTs (Pang et al, 2005;Davis et al, 1986), but the GC content of the third position of the codon in this gene is very high. The possible reason is the high evolution pressure to the CaM gene (Griess et al, 1989(Griess et al, , 1994Xu et al, 2006). The context of the translation initiation codon in CaM gene sequences is GCGCCAUGCG, and it is highly similar to the Kozak consensus sequence CC (A/G) CC AUGGC (Kozak, 1987).…”

Section: Characteristics Of the Nucleic Acid And Deducedmentioning

confidence: 98%

Cloning and analysis of calmodulin gene from Porphyra yezoensis Ueda (Bangiales, Rhodophyta)

Wang

Mao

Zhuang

et al. 2009

J. Ocean Univ. China

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In order to understand the mechanisms of signal transduction and anti-desiccation mechanisms of Porphyra yezoensis, cDNA and its genomic sequence of Calmodulin gene (CaM) was cloned by the technique of polymerase chain reaction (PCR) based on the analysis of P. yezoensis ESTs from dbEST database. The result shows that the full-length cDNA of CaM consists of 603 bps including an ORF encoding for 151 amino acids and a terminate codon UGA, while the length of genomic sequence is 1231 bps including 2 exons and 1 intron. The average GC content of the coding region is 58.77%, while the GC content of the third position of this gene is as high as 82.23%. Four Ca 2+ binding sites (EF-hand) are found in this gene. The predicted molecular mass of the deduced peptide is 16688.72 Da and the pI is 4.222. By aligning with known CaM genes, the similarity of CaM gene sequence with homologous genes in Chlamydomonas incerta and Chlamydomonas reinhardtii is 72.7% and 72.2% respectively, and the similarity of the deduced amino acid sequence of CaM gene with homologous genes in C. incerta and C. reinhardtii are both 71.5%. This is the first report on CaM from a species of Rhodophyta.

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“…A calmodulin cDNA isolated from roots of maize inbred B73Ht seedlings (Ref. 18, gift from Dr. G. Feix, Germany) was PCR-amplified and subcloned into pQE-30. The clone was overexpressed, and the recombinant CaM was purified to homogeneity using the QiaExpress system.…”

Section: Expression Of Cap1 Product In Yeast and Complementation Of Ymentioning

confidence: 99%