We present the results of analysis of membrane phosphoproteomes from individual morphological stages of Streptomyces coelicolor that reflect developmentally dependent heterogeneity and phosphorylation of intrinsic and externally added purified Strepomyces aureofaciens EF-Tu. Fast growing nonpathogenic Mycobacterium smegmatis was used as a non-differentiating actinomycetes comparative model. Streptomycetes membrane fraction was found to contain protein kinase(s) catalyzing phosphorylation of both its own and an externally added EF-Tu, whereas Mycobacterium membrane fraction contains protein kinase phosphorylating only its own EF-Tu.

• MeSH
• buněčná membrána chemie   enzymologie   metabolismus   MeSH
• elongační faktor Tu izolace a purifikace   metabolismus   MeSH
• fosforylace MeSH
• Mycobacterium smegmatis chemie   enzymologie   metabolismus   MeSH
• posttranslační úpravy proteinů * MeSH
• proteinkinasy izolace a purifikace   metabolismus   MeSH
• Streptomyces chemie   enzymologie   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• elongační faktor Tu MeSH
• proteinkinasy MeSH

In vitro phosphorylation of EF-Tu was shown in cell-free extract from dormant spores of Streptomyces coelicolor by a protein kinase present in spores. EF-Tu phosphorylation was observed on both intrinsic S. coelicolor factor and externally added purified EF-Tu from S. aureofaciens, on two isoforms. Putative serine and threonine residues as potential phosphorylation targets were determined in primary sequence and demonstrated on 3D structure model of EF-Tu.

• MeSH
• 2D gelová elektroforéza MeSH
• elongační faktor Tu chemie   izolace a purifikace   metabolismus   MeSH
• fosforylace MeSH
• molekulární modely MeSH
• molekulární sekvence - údaje MeSH
• proteinkinasy metabolismus   MeSH
• sekvence aminokyselin MeSH
• sekvenční seřazení MeSH
• spory bakteriální enzymologie   metabolismus   MeSH
• Streptomyces aureofaciens metabolismus   MeSH
• Streptomyces coelicolor metabolismus   MeSH
• terciární struktura proteinů MeSH
• zobrazování trojrozměrné MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• elongační faktor Tu MeSH
• proteinkinasy MeSH

Some molecular properties of the elongation factor Tu of protein synthesis purified in an aggregated state from gram-positive Streptomyces aureofaciens were studied and compared with those of Tu from gram-negative Escherichia coli. Electrofocussing under reducing conditions showed that the molecule of EF-Tu from S. aureofaciens has an isoelectric point shifted more to the acidic side compared with EF-Tu from E. coli. A comparison of amino acid composition revealed minor differences in the content of several amino acids in the two factors and showed that EF-Tu from S. aureofaciens contains four half-cystines per molecule. Under denaturing conditions only two mercapto groups reacted with 5,5'-dithiobis(2-nitrobenzoic acid). Limited tryptic digestion of aggregated EF-Tu from S. aureofaciens yields six fragments: the four main fragments are of a similar size as those of the E. coli factor. All fragments detected after trypsin digestion of S. aureofaciens EF-Tu were immunologically cross-reactive with antibodies against E. coli EF-Tu. However, even after 2 h of the reaction there still remains a small part of streptomycete factor uncleaved, which documents high resistance of aggregated EF-Tu towards trypsin.

• MeSH
• aminokyseliny analýza   MeSH
• elektroforéza MeSH
• elongační faktor Tu izolace a purifikace   MeSH
• Escherichia coli analýza   MeSH
• hydrolýza MeSH
• isoelektrická fokusace MeSH
• Streptomyces aureofaciens analýza   MeSH
• sulfhydrylové sloučeniny MeSH
• trypsin MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• aminokyseliny MeSH
• elongační faktor Tu MeSH
• sulfhydrylové sloučeniny MeSH
• trypsin MeSH