It’s a lot harder to detemine structure without being able to get crystals. you need to be more clever.
Computer modeling and force fields are very useful tool now. Back when you were doing that they quite nascent and needed expensive computers.
Did you ever figure out why your protein was so unstable?
Biologically, we had a very good reason for the instability of the protein.
Chemically, I was never able to show anything definitive. I loaded up my reactions with protease inhibitors, but the protein showed the same degradation pattern on gels. There was always the intact protein band at the top (about 95 kDa), another major band at about 50 kDa, a third major band at about 30 kDa, and between the major bands were several minor bands/smear.
I know that the cDNA had some cryptic Kozak sequences, which could explain the two smaller major bands. But they did not account for the minor bands/smear.
If I were to make a standard control (from liver extracts) for Western blotting, and store it ready to load on a gel in the -20, it would degrade within a couple of weeks. That was after it was mixed with the beta-mercaptoethanol/ SDS loading buffer and boiled for 2 minutes. The raw extracts and in vitro synthesized proteins had to be stored at -80.
Needless to say, for structural studies, this protein was quite frustrating. However, I was able to generate some very nice functional and protein interaction data, sufficient to earn a PhD, and I can’t complain about that.
I have not worked with that protein since graduate school, and I don’t know if there is any structure data other than computer models.
I must say, it is unusual to encounter someone on FR who understands the language of biochemistry. Most of my PhD scientist colleagues at work don’t even understand it. :)