Research on human vascular proteins

No amount of experimentation can ever prove me right; but a single experiment can prove me wrong - Albert Einstein

Long way since the black box concept
In the '70s molecular cell biology was still in its infancy!

A bit like imagining life without the smartphone, it's hard for graduates entering life sciences today to imagine research with no genomic data, little available on the fledgling internet, no world wide web, no real understanding of molecular cell biology and literature research done in the library (most often through a discussion with a trained librarian a.k.a. information professional) and by sending reprint requests. That was the situation when I started postgraduate research at Oxford in 1970.

Useful Internet sites like Pub Med and apps like Research Gate have rendered the days of taking journals off library shelves redundant and research workers don't really have to leave their labs. BUT a note of caution has to be sounded here. In one of the most productive life science research labs in the world; the Laboratory of Molecular Biology in Cambridge (UK!) its director in those days - one Max Perutz - absolutely insisted researchers left the lab and communed in the coffee area. The 'cross fertilisation' of ideas that resulted speaks for itself. At the MRC National Institute for Medical Research (the largest of the MRC Units), the divisions effectively kept themselves to themselves and this valuable "meeting of minds" didn't happen. Not just fundamental research institutes; but also industry: please note!

Looking back now in 2021, my research career has embraced human vascular biology in general, although there was certainly no intention to do that at any particular time. At Oxford I was painfully able to solve the molecular structure of the human plasma thyroid hormone transporter protein: transthyretin (TTR; but it was known as prealbumin then). Only one other plasma protein 3D structure (IgG) had been solved at that time. Exhausted by the effort, I spent two years teaching at Hertfordshire University then took up a post at the MRC's National Institute for Medical Research in Mill Hill, London.

My boss there was interested in whole blood structure and function and I was set to look at the physical and binding properties of human serum albumin (HSA). Although I was a biophysicist by background I was happy to teach myself some real biochemistry. This was laborious in those days by comparison with the sophisticated and time-saving automated instrumentation available now. But I was able to separate HSA into large domains each retaining specific binding properties. This experience was to prove very useful years later in a biotech company where I was able to ask the molecular biologists to construct cDNAs to express HSA domains instead of using proteases to cut the intact protein!

Human serum transthyretin structure. The dimer illustrated is half the molecule: the whole protein is a tetramer of identical subunits

Annexin core structure

After making my boss (the head of Biophysics department) happy with the HSA work, I toured the institute: a molecular Maverick, looking for projects with the potential to get me a permanent appointment (a.k.a. tenure). I had an exciting year with Derek Smythe in the Peptide Chemistry Division working on endorphins when the news about the analgesic activities of these pituitary peptides broke. A flurry of papers was published. I also have a cautionary tale about talking to the press about your research! After working on endorphins, interested in the release of hormones into the circulation, I decided to investigate the molecular mechanism of hormone secretion (a.k.a. exocytosis). If I had had a mentor (always a good idea if any postgraduates or post-docs are reading this) he or she should have gently steered me away from such an ambitious project; one certainly requiring multidisciplinary teamwork. I should at least have been a card-carrying cell biologist rather than a lone biophysicist. Long story short, this led to my recognition of a new family of calcium-dependent membrane binding proteins which I christened: Annexins.

Interesting as the annexins were, there was at that time no clear cell functions supported by these proteins (35 yrs later they are still rather enigmatic) and I had a tenure to gain. Fascinated by the whole process of intracellular traffic (plasma membrane recovery is an important component of exocytosis) I began to study the key mechanisms of endo- and phago-cytosis. This time I had learnt an important lesson and my later work at Mill Hill was more collaborative. I was able to monitor the environment of bacteria taken up by human neutrophil phagosomes and follow receptor-mediated uptake in cell culture.

A lot of ground-breaking research happened at the MRC's National Institute for Medical Research. Now its own ground is broken and it has become a block of flats! Staff moved to the Crick Institute in London.

Albumedix recombinant HSA

A new Institute director took over the reins at NIMR and, although tenured staff by then , I decided that I should pursue my career in industry as a protein scientist. I joined Delta Biotechnology and found myself - once again- studying HSA, this time as a recombinant product. At that time HSA was widely used in clinical medicine and was a fraction separated from donor blood plasma. There were serious concerns over the actual and potential transmission of virus (HIV and hepatitis) from this source and recombinant HSA was regarded as a safer option (around 12 gms was typically infused into patients). Delta was successful in devising a very efficient producer organism for recombinant HSA, quality control and downstream processing. I left the company when its owners put it up for sale, but its successor, ALBUMEDIX, has brought recombinant HSA to the market.