# In Vivo Ribosome-Amplified MetaBOlism, RAMBO, Effect Observed by Real Time Pulse Chase, RTPC, NMR Spectroscopy

## TL;DR

Scientists confirmed a process called RAMBO, where ribosomes influence metabolism in living cells, using a new noninvasive method.

## Contribution

The RAMBO effect was validated in vivo for the first time using real-time pulse chase NMR spectroscopy.

## Key findings

- The ribosome-amplified metabolism effect was confirmed in living Escherichia coli cells.
- Chloramphenicol disrupted pyruvate kinase's quinary structure and inhibited the RAMBO effect in vivo.
- The RTPC-NMR platform enables high-resolution metabolic monitoring in real time across cell types.

## Abstract

Quinary interactions
between proteins and ribosomes play
an important
role in regulating biological activity through a phenomenon termed
the Ribosome-Amplified MetaBOlism, RAMBO, effect. This effect has
been documented in vitro but not in vivo. Real time pulse chase, RTPC,
NMR spectroscopy, coupled with isotopic flux analysis in Escherichia coli was used to validate the RAMBO effect
in vivo. The ribosomal-targeting antibiotic chloramphenicol was employed
to disrupt the quinary structure of pyruvate kinase, the final enzyme
in glycolysis. Kinetic flux profiling demonstrated that the in vitro
deactivation of the RAMBO effect by chloramphenicol was also observed
in vivo, thereby confirming the potential role of ribosomes in regulating
glycolysis. The noninvasive modular design of the RTPC-NMR platform
allows for high-resolution metabolic monitoring across different cell
types, providing broad applicability for studying the real-time metabolic
responses to external stimuli in living cells.